Learning Objectives 1. Define and use correctly the terms aldose, ketose, pentose, Recognize the structures of glucose, fructose, glucuronic acid, glucosamine, and N-acetyiglucosamine. Explain the o, . nomenclature system for simple carbohydrates; note that it does not predict the direction of rotation of plane polarized light. Explain what is meant by the term reducing sugar. Distinguish between the reducing and nonreducing ends of a polysaccharide such as glycogen. List several common biologically relevant modifications of simple sugars. List three common disaccharides and their natural sources. Describe how they ~ differ in structure. Describe the structure of common simple polysaccharides such as starch, cellulose, and glycogen. List several common glycoproteins and give their function, Explain how sugars are linked to proteins via asparagine and serine or threonine. Describe differences in the glycolipids, especially the carbohydrate content, contributing to the different blood ‘rOUp artigens. Explain the locher'ieal basis for these differences. esciba on sletes Doo! suost cen ined iain Goueyiaar ane how quantitation of HbA, is used in following the effectiveneas of reatment of diabetes. Relate the release of heparan sulfate after tissue injury to its effects on blood clotting. Identify the differences between heparan sulfate and heparin. 2122 Chapter 2 Carbohydrates. Simple Sugars: Monosaccharides Introduction Carbohydrates (sugars and polymers made of sugars) are ubiquitous in biology and play many important roles. Here are just a few of the many ways carbohydrates are basic to biology: © The simple sugar glucose is a principal source of metabolic energy for the cell. Cellulose,a polymer of glucose, strengthens the cell walls of plants. * The cell uses carbohydrate chains to modify proteins and direct those proteins to their proper sites for function in the cell. * In animals, the complex polymers known as glycosaminoglycans (chains of unmodified and modified sugars) lubricate and cushion joints; help form skin, hair, and feathers; help regulate blood clotting and assist the immune system; and help control cell-to-cell adhesion and interaction. Basic Nomenclature The formula for a carbohydrate, (CH;O).. may be simple, but there is a great deal of complexity in the structures of such compounds. Inevitably, to properly describe this structural diversity, there is also a great deal of detailed nomenclature. Conventionally, a monosaccharide is a carbohydrate that does not hydrolyze, while a disaccharide can be hydrolyzed to give two monosaccharides, a trisaccharide can be hydrolyzed to give three monosaccharides, and so on. A polysaccharide contains many monosaccharide units. Also, the smallest molecules regarded as carbohydrates have three carbon atoms; the exemplars here are glyceraldehyde and dihydroxyacetone (Figure 2-1). Compounds with only ‘one or two carbons do not share extensive properties with those we regard as “sugars,” so they are not regarded as saccharides, “The suffix'"-ose” on a compound’s name indicates a sugar. An aldose is a sugar derived from an aldehyde, and a ketose is derived from a ketone. A triase is a carbohydrate with three carbon atoms; a tetrose has four carbons; a pentose has five carbons; and a hexose has six carbons. These terms may be combined for a more complete specification of the compound. For example, an aldopentose is a sugar with five carbons and an aldehyde group. Stereochemistry ‘A very important feature of sugars is the large number of isomers possible within the overall gmolecular formula, even for simple pentoses and hexoses.A. quick review of some concepts and terminology from organic chemistry will help to keep matters clear. ‘© Stereoisomers are molecules that have the same molecular formula but differ in the way that the constituent atoms are oriented in space. Figure 2-1 The smallest saccharides, glyceraldchyde and dihydroxyacetonc.‘Simple Sugars: Monosaccharides 23 ‘¢ Enantiomers are molecules that contain a center of asymmetry, usually a carbon atom known, asa chiral (“handed”) carbon. Enantiomers are mirror images of each other. They have the same physical properties, except they rotate polarized light in different directions. By convention, a + sign indicates rotation of the polarization to the right, while a ~ sign indicates rotation to the left. Because their physical properties are the same, it is often very difficult to physically separate enantiomers from each other. + Diastercomers are stereoisomers that are not mirror images. Their physical properties differ, and this permits their physical separation. Note that a pair of stereoisomers that are not enantiomers must then be diasteromers. Glyceraldehyde contains a chiral carbon, and there are two possible enantiomers of this compound. Figure 2-2 illustrates these two forms using a Fischer projection. By convention, the C1 carbon is the aldehydic carbon, and it is drawn at the top of a vertical line of carbon atoms, C1 through C3. The chiral carbon is C2; the OH group is drawn to the right of the chiral carbon for what is denoted as the D isomer; the L isomer has this group drawn to the left side of this carbon. In terms of visualizing the direction of bonds around C2, the vertically aligned bonds are imagined to be going into the plane of the drawing and away from the viewer, while the horizontally aligned bonds are directed upward, out of the plane of the drawing and toward the viewer. Figure 2-3 presents the same structures using wedges in place of lines, to give a better sense of perspective on the direction of the bonds. ‘With respect to rotation of plane polarized light, the 1» isomer here rotates it to the right (che p originally indicated dextro, or right-handed, rotation and the 1 indicated fevo, or left- handed, rotation). The D, L convention for stereoisomers can also be applied to other simple chiral compounds, such as amino acids. However, while many » monosaccharides do, indeed, rotate plane-polarized light to the right, this behavior is not universal. Instead, to indicate thi particular physical property, as distinct from the configuration about the chiral carbon, it is preferable to use a prefix of + or —, with the + sign indicating right-handed rotation and the — sign indicating left-handed rotation. Figure 2-2 p- and t-glyceraldehyde as Fischer projections. D-Giyceraidenyde L-Glycoraldehyde Figure 2-3. p- and t-glyceraldehyde drawn using “wedge” bonds.7 2 Carbohydrates bo 20 HL 20 Ne, a, ¢ Ses THo-c— —C—H. CoH cHont Home Hoey pe EH H-¢0H ad cH,0H GH,0H SEnthose ——LErythose p-Tmeose Flgure 2-4 Pairs of enamiomers forthe adoteoses - and Leerythrose, and D- and t-thre Tetroses have four carbons, ewo of which may be chiral. Consider first the Possibilities gy the aldotetroses. Figure 2-4 shows erythrose and threose; there in two PEL DaeES for each & these compounds, so there are four stereoisomers overall—that her osha Of emotionen Note that either enantiomer of erythrose is a diasteromer of either aniiomnes of Either threose or erythrose can be tautomerized to a ketose, erythulose; ee PTOI ging D-erythulose, which in turn gives D-threose (Figure 2-5). A similar pattern holds fo. the interconversion of the L isomers. , With aldopentoses there are three chiral carbons, giving four at pairs of enantiomer, D and t forms for ribose, arabinose, xylose, and lyxose (Figure > D ioe fe Ketopentoses, ribulose and xylulose, each with aD and an L form. The D isomers are ie HL 0 ¢ Wo=G-H H—d—on bon o-Tveose Pentoses. Only the » isomers are shown, Figure 2-6Simple Sugars: Monosaccharides 25 most important for biochemistry. Additionally, the important pentose derived from ribose, deoxyribose, is essential to the structure of DNA. There are 4 chiral carbons for aldohexoses and 16 diastereomers. b-Glucose, D-mannose, and p-galactose are the most important for human biochemistry. There are also four ketohexoses, with D-fructose being the most important species. D-(+)-Glucose is sometimes referred to as dextrose, while levulose is b-(—)-fructose. Cyclization The aldopentoses and the hexoses show a strong tendency to cyclize in solution, forming rings with five or six members. These rings contain an oxygen atom as one of the members. Thus sugars with a five-membered ring resemble the compound furan, while those with 2 six-membered ring resemble pyran (Figure 2-7). This leads to the practice of designating the sugars in such conformations as furanoses and pyranoses, respectively. As a pyranose, glucose forms a hemiacetal (the product of a 1:1 reaction of an aldehyde with an alcohol; see Figure 2-8), which is relatively stable thanks to its cyclic structure. This reaction creates a new chiral center at C1 and leads to two new optically active forms, called anomers; C1 is referred to as the anomeric carbon in this case. The anomers are designated as «-p-glucopyranose and B-b-glucopyranose (Figure 2-9); these names are often abbreviated simply to a-p-glucose and B-p-glucose. Figure 2-9 shows these anomers in the stable “chair” conformation. Here the —OH groups at C2, C3, C4, and C5 are equatorial. However, notice that the — OH group at C1 may be either equatorial (the B anomer) or axial (the o anomer). aed gO wv Figure 2-7 Furan and pyran. P pe RK + RCH ——— A—Choc yyy Hemiaceal iH 4 ou f+ meHoH ——~ aoc. ae LOH Soot, Hemiketal a Figure 2-8 Hemiacetal and hemiketal formation. iy pose ty yen ase 4 HON Leg rN Ta @-0-Glucopyranose B-0-Glucopyranose Figure 2-9 Anomers of p-glucose. Some bond lengths have been extended to emphasized structural details and the — OH group attached to the anomeric carbon has been marked.28 Chapter 2. Carbohydrates ¢, through formation of , * Fructose can cyclize to give either a furanose ring ic ‘bony! of a ketone). For hemiketal (the product of a nucleophilic alcohol reacting ible, et- and B-D-fructofuranose either the pyranose or the furanose, two anomers are possible, @= AN" ae and a- and B-p-fructopyranose (Figure 2-10). In solution, te Pe tc anioute of (approximately 60%) over the furanose form (approximately (Figure 2-11) can also cyclize open-chain form (much less than 1%). Ribose and deoxyril to form a furanose, with two anomers. Reactivity and Common Derivatives ‘The hemiacetals and hemiketals formed in cyclizing pentoses an erally favors the cyclized with the open-chain form of these sugars, although the equilibrium Bo°° apes there will be form by a considerable margin. As a consequence, in solutions of Sess MEANT | A TOs smnall amounts of “fee” aldehyde or Ketone available for reaction: MURINE TT can be oxidized by mild agents. This behavior is the basis fee Se car ntagent or de cole lab tess for an aldehyde, such asthe silver tnirror produced by Tolens’Teazis Ot XX Solos changes produced by Benedict’ solution or Fehling’ solution. Because solu © 2 CS nd ketoses will have small amounts ofthe “ee” aldehyde or ketone, they can #0 tt XA ee Feagents. Sugars that give positive test results with these reagents are te 7 ‘The products of the oxidation of aldoses are called aldonic acids. esand hexoses are still in equilibrium Figure 2-10 Anomers of v-fructose as a pyranose and faranose. GOH OH SHO On 4 ¥ - " HK # A, . OW >-Ribose on Figure 2-11 p-Ribose and p-deoxyribose. és,‘Simple Sugars: Monosaccharides 27 Conversely, the carbonyl group on aldoses and ketoses may be reduced to give the corresponding alcohol. Figure 2-12 shows two important examples of this class of compounds. p-Sorbitol is also called p-glucitol. In the disease of diabetes mellitus, sorbitol accumulates in the lens of the eye and is associated with the formation of cataracts. The mechanism of cataract formation may possibly involve redox reactions in the formation of sorbitol from glucose and associated oxidative cross-linking of proteins through disulfide bridges. Enzymes can oxidize monosaccharides to give a number of different products. Two important carboxylic acids that are derived in this way are glucuronic aid and iduronic acid (Figure 2-13).Note that iduronic acid is an epimer of glucuronic acid; an epimer is one of a pair of diasteromeric aldoses that differ only at the C2 position in their configuration. CGilyesides are formed from hemiacetals or hemiketals (that is, reducing sugars) by reaction with an alcohol. The product is a fall acetal or full ketal (Figure 2-14). Glycosides are not reducing sugars. In naming these compounds, “glyc” is the general prefix for the combination oH0H oH.OH H—G—o4 Ho-¢—H Ho— G4 Ho-¢—H H—-G-04 H—G-0H ty ee 0H HOH o-Soritl o-Mannitot Figure 2-12 Sorbitol and mannitol are cwo important alditols, derived from the reduction of the corresponding aldoses. Q %e » ATR ‘OH CGlucuronie Acid uronic Acid Figure 2-13 Uronic acids: glucuronic and iduronic acid. we ae eee eu @-D-Glucose * Alkylglucoside: CHO OH OR + ROH tne CH,OH ‘CHZOH On OH -D-Fructoturanose: Alkyttructoturanoside Figure 2-14 Glycoside formation, giving a fall acetal or fll ketal.2B Chapter 2 Carbohydrates He w-6-0n q brotd ki A PK, = 0.97 arti rent pee me e ope 4 ; HO: HO: p HO, a o OH wv 1-6-phosphate wi eneose mee pi 094 pobre pie = 64 Figure 2-15 Sugar-phosphate esters and theit pK, values. of an unspecified sugar and alcohol. Thus reaction with methanol would give a methylglycosde, for example. If the sugar is specifically identified, one may have 2 glucoside (from glucose), a galactoside (from galactose), and so on. Glycosides are fairly stable and are not readily cleaved by hydroxide ion. However, the bond linking the alcohol and sugar can be cleaved by enzymes called glycosidases. Glycosides are widely distributed in plants and animals. "As alcohols, sugars can react with phosphoric acid to form phosphate esters. These compounds are quite important in the metabolism of carbohydrates, forming many important intermediates. Sugar-phosphate esters are acidic, even more so than orthophosphoric acid. Figure 2-15 collects several important examples of carbohydrate phosphate esters that are important in energy metabolism. Hydrolysis of these compounds is thermodynamically favorable, a point that we will return to later (Chapter 9) as we discuss carbohydrate metabolism. ‘Natural polysaccharides may contain sugars that have been modified by sulfation or attachment of an amino group. Two common amino sugars are B-p-glucosamine and B-p-galactosamine (Figure 2-16). These amino sugars can be further modified by acetylation ‘or other alterations. Also, in these natural polysaccharides, sulfate groups may be attached to sugars in a variety of ways (Figure 2-17). When dealing with these more complex compounds, it is convenient to use abbreviations for the monosaccharides and their derivatives. Table 2-1 collects some of the most commonly used abbreviations. Oligosaccharides and Polysaccharides “The terms “oligomer” and “polymer” are used to indicate : | seeimple consGanion. The siople units are Called monomers seepienacepa oflinked Fed Slonoraers is called a polymer. An oligomer is eseentally a short polymers it con ining many more monomer units, but not as many as a polymer. The size hier it contail me and an oligomer is not well defined. It is safe to say that a chain of 100 sentir t ra wouldFigure 2-16 Amino sugars and their derivatives. 2-N-Sullonato-3,6-bie-O-sullonatoglucosamine Figure 2-17 Sulfated sugars found in natural polysaccharides.89 Chapter2 Carbohydrates wea Meet ee Monosaccharide Abbreviation Glucose Glc Fructose Fru Galactose Gal Mannose Man Xylose xyl Fucose Fuc B-p-acetylgalactosamine ___GalNAc B-p-acetylglucosamine GleNAc Glucuronic acid GleA Iduronic acid IdoA Sialic acid Sia = Stabiecid 20 Si Table 2-1 Common Simple and Modified Monosaccharides Found in Glycosaminoglycans qualify as 2 polymer, and a chain of only 10 or 20 units would probably be classified as an oligomer, but a chain of 50 units might in some cases be referred to as a polymer, and in other instances as an oligomer. The prefixes “poly-""and “oligo-""can be combined with the name of the type of monomeric unit to specify the type of polymer or oligomer. For example, a long chain whose repeating monomeric units are all sugars would be regarded as a polysaccharide; a short chain of two or three nucleic acid units (nucleotides) might called an oligonucleotide, and so on. Oligosaccharides There are three common disaccharides: sucrose, lactose, and maltose. Sucrose, or common table sugar, is commercially prepared from sugar cane or sugar beets. It is widely found in plants, Sucrose itself is not a reducing sugar, so there are no “free” hemiacetal or hemiketal moieties in the disaccharide. Upon hydrolysis, sucrose gives D-(-+)-glucose and p-(—)-fructose. ‘These monomers are linked through the anomeric’ carbons, C1 of glucose and C2 of fructose (we shall abbreviate this as a 1— 2 linkage), as shown in Figure 2-18, to form «-p-glucopyranosyl-p-p-fructofuranoside, which is both a glucoside and a fructoside. Figure 2-18 Sucrose: a-p-glucopyranosyl-B-p-fructofuranoside.Oligosaccharides and Polysaccharides 31 os Sita ares, Z ‘ ie ee fs on Lactose (b Anomer) Figure 2-19 B-Lactose: 4-B-p-galactopyranosyl-p-glucopyranose Lactose is present in milk at approximately 5% concentration and is commercially prepared asa dairy by-product. A disaccharide of galactose and glucose, itis itself a reducing sugar. with the glucose attached to one of the oxygens of the galactose unit. This makes lactose technically 2 galactoside, not a glucoside; it is the glucose unit with its “free” hemiacetal that gives the disaccharide its character as a reducing sugar. Formally, lactose is 4-B-p-galactopyranosy!- p-glucose (Figure 2-19). The breakdown of starch yields maltose. This disaccharide is a reducing sugar, which can be hydrolyzed to two molecules of glucose. The linkage is through an acetal, and there are (wo possible configurations of the anomeric carbon involved: a-maltose (4-c-glucopyranosyl-a “p-glucopyranose) or B-maltose (4-a-glucopyranosyl-B-p-glucopyranose). See Figure 2-20. In later sections and chapters, we will usually drop the detailed specification of configuration and simply assume the configurations listed here. Glucose Polymers Starch is a plant storage form for glucose; it is a polymer of glucose. Acid hydrolysis converts starch to corn syrup. There are two main fractions: water-soluble amylose (about 20%) and water-insoluble amylopectin (about 80%). Amylose is a linear polymer of around 200 units of glucose with a (1 — 4) linkages (Figure 2-21). Amylopectin is also a glucose polymer, but of z my errs Mattose (a: Anomer) Figure 2-20 a-Maltose: 4-cr-glucopyranoxyl-c.-b-glucopyranose. con,ou ? XS 3 7 6 wi bs) HOH Ho. ‘OH O-4 Figure 2-21 Amylose. Hydrogen atoms have been omitted for clarity.32 Chapter 2. Carbohycrates motecule), with occasional brag, ie higher molecular weight (at least 1000 elvcos units per 4 linkages. but the branche, in the chain; most of the glucose residues a joined by a formed using ce(1 —> 6) linkages. «tar eo amy Glycogen is an animal polymer of glucose, similar £0 7" Chapter 9- Cellulose is the mr Its synthesis and breakdown are the subject of discussion 17 Tp is insoluble in water wn polymer of the cell walls of plants: it forms woody structures T 1, per molecule. Al bovtt starch and glycogen. In cellulose, there are a few chousand sINCOS most animals The thou, flan in ghavose polymer, iis not igesible PY humans Oh (Figre 2-22), and bre tits are joined by B(1 —> 4) linkages, unlike glycogen oF ponds. Ruminant cin generally do not make the enzyme needed (0 hydrolyze these Bones ceria that make qe Beer and inseces such as termites carry in thelr BM certain oats to digest cel shane te enzymes necessary to break down cellulose, enabling their animal HOY oe forri tose. ‘As a part of the cell wallthe polymer chains of cellulose lie 5 ee Facsd coats bundle hese burtdles are ewisted together to create sands and finaly visible bet contains lignin (Cros-linked C9 aromatic units, related in stracte® ed bispathesist the aromatic amino Figure 2-23) embedded in cellulose fibers the lignin gives the wood Very igh ene Glycosaminoglycans : ; Glyeosaminoglycans (GAGs) are large linear polymers of repeating disaccharide units, commonly containing one or another amino sugar 2% cone of th the disaccharide unit. Several types of glycosaminoglycans exist including chondroitin ; versean salfate:they differ in the ype of disaccharide anit used in their synthesis. Glycosaminogicars are found outside cells, on the cel surface, oF a5 Part “ofthe extracellular matrix. They perform ae ea jan serving as mechanical support and cushioning joines, as cellular signals involved in {all proliferation and cell migration, and as inhibitors of certain key enzymes. Giycosaminoglycans are offen found attached to 2 Pru core, forming what is called a proteoglycan. inthe past proteoglycans were called mucopolysaccharides. Proveoglycans are more carbohydrate than protein, £0 their properties are mainly determined by the carbohydrate portion of the molecule, These carbohydrate moieties may contin os. cae oxen " wT f ma OH Figure 2-22 Cellulose. Hydrogen atoms have been omitted for clarity. pectin, but with more bran Figure 2-23 Lignin (partial structure).Oligosaccharides and Polysaccharides 33 carboxylic acids (uronic acids) or sulfated sugars, so generally the GAG chains carry a substantial negative charge. Charge repulsions will tend to drive the polysaccharide chains into an extended conformation. Additionally, the charges will help to attract water molecules, so the chains are highly hydrated. The typical proteoglycan is formed from multiple GAG chains on a single polypeptide chain. The overall structure might be described as “feathery”; see Figure 2-24. Heparan sulfate (HS) is a sulfated polysaccharide, found as a component of cell-surface proteoglycans in mast cells, and on the surface of endothelial cells lining blood vessels. It is composed of repeating units of N-acetylglucosamine and uronic acids (either glucuronic or iduronic acids). Sulfation (sulfate ester formation) can be found at several positions on these residues; also, the acetyl group on N-acetylglucosamine may be replaced by a sulfate group (Figure 2-25). After an injury to tissue, oligosaccharides derived from this GAG are released and subsequently help to mediate the inflammatory response. Some of the released oligomers Figure 2-24 Proteoglycan structure. Core protein strands are heavily modified with keratin sulfate and chondroitin sulfate. The core protein strands are held in a complex with a strand of hyaluronic acid by link proteins. 4 HO Ho Osos oon t He Ts m food? NH 8 HL come 2-O.Sullonatoiduronic acid 2-N'Sulfoneto-6-Osulfonato-glucosamine Figure 2-25 Heparan sulfate.34 Chapter 2 Carbohydrates Ss icoiti ‘ Promote activity by growth factors, chemokines, and icant the action ofa Fecruitment of leukocytes to the injury site. A very importazrOGthely | ga Paticu Pentasaccharide sequence as an anticoagulant. tee ate as heparin. The pentasacet : : —— : esi pabomers with this pentisaccharide sequence If TPM zyme is responsible for inhign* binds to and activates the enzyme antithro: err i thrombin;a protsare eae forming blood clots;this inhibition of thrombin, in turn, bigs blood clotting. Note that heparin is much smaller than heparan ae li to a protein core. Heparin is also more highly sulfated than the Eee ey aacchan; ide Sequence in heparan sulfate. synthetic version of the key pentasite WNICE TAS ctnica an anticoagulant (see the discussion later in this chapter in connection With Figure 2-35), Chondroitin sulfate (CS) is a relatively shore polymen conststg O | 1 AINE, Pesiducs op glucuronic acid and galactose N-acetyl 4-sulfonate (Figure ‘ a ‘Neacetyl ulfate (Ds) i, a closely related GAG, which is composed of glucuronic acid ani Eee = igalactosaming. Chondroitin sulfate is found in the extracellular matrix. ts roles are mainly ss lend mechanicy, support and flexibility to tissue; notably, it helps to form skin and cartilage. On membranes, cg, and DS are involved in interactions with receptors for growth factors, where they may serve cofactors for various growth factors. : Pohick Keratan sulfate (KS) is found in proteoglycans in three forms, two of which are branched KS is found primarily in the cornea of the eye and in joint cartilage, where it serves mainly in a mechanical/struetural role. It is formed from alternating units of galactose and sulfated N-acetylglucosamine (Figure 2-27) : Hyaluronic acid (HA; also called hyaluronate) is a long, linear polymer of alternating N-acetylglucosamine and glucuronic acid residues (Figure 2-28). It serves as a lubricant in joints in the form of synovial fluid; HA is also a principal constituent of the vitreous humor in the eye, 0,80 on K 4 OF my ways 4 Hon 4 \ Hy Glucuronic Acid 2-N-Acetyi-4-O-sulfonato-galactosamine Figure 2-26 Chondroitin sulfate. onoH 4 poet te Ho ee Ho. One 4 4 Hoed Galactose Hs “ &-O-Sutfonato-2-Nacetyighucosamine Figure 2-27 Keratan sulfate.Glucuronic Acid ‘N-Acetyiglucosamine Figure 2-28 Hyaluronic acid - and it helps to form cartilage. Unlike other GAGs, this polymer is neither sulfated nor attached to a protein core. It is secreted directly to the extracellular matrix. The degradation of HA that occurs after tissue injury releases smaller chains, which can participate in cell proliferation, migration, and differentiation; these degradation products also help to recruit leukocytes to the site of injury. Intra-articular injections of purified HAA have been used therapeutically in cases of osteoarthritis. Other Natural Polysaccharides of Interest ‘Agar isa linear polymer of sulfated and unsulfated galactose residues, prepared from marine algae; agarose (Figure 2-29) is derived from agar as the mostly unsulfared fraction. It is an alternating copolymer of galactose and 3,6-anhydro-galactose. This polymer is not 2 proteoglycan, but rather is purely carbohydrate. When dissolved in hot water and then cooled, it forms gels with suitable strength for lab use in electrophoresis; itis also used as a food additive to thicken liquid suspensions. Chitin (Figure 2-29) is a polymer of N-acetylglucosamine units. The linkage between units js similar to that in cellulose; the main structural difference is that the C2 carbon now bears ‘an amino group that is acetylated, instead of a hydroxyl group. Like cellulose, chitin is used for structural purposes; it forms the hard exoskeleton of arthropods (¢.g., insects, spiders, and crabs). bho os ok * ‘Agarose oe CH o=d cHyoH ‘NH CH,OH . pe Nat Pe HO. O- HO. O- Sa as ot i { { CH Hy of Figure 2-29 Agarose and chitin. Note the unusual ether bridge on the sulfited anhydrogalactose unit in agarose.96 Chapter 2 Carbohydrates Biosynthesis of Proteoglycans ee ‘of mammalian proteoglycans lacing attachment of a xylose residue to the side chain o| ices (Flesselz. residues and a ghicuronie acid residue are then attached oe erent euayrics capt pathways leading to the different GAGs diverge at this Poeaifing the residues afer thet the joining of the sugar residues and more enzymes MOCIIOK "Oy sain elongation joined. Chain initiation takes place in the endoplasmic recicwlO™ 27° Cy’ eas modification occur in the Golgi complex. The proteosivcane NON’ ari biog the extracellular matrix, carried by vesicles that bud off d m6 £6 is not attached toa pros’ path is not applicable to hyaluronic acid, however, because (is Protein but is instead extruded directly to the extracellular matrix and the associated GAG: 2S ih fa gerine on the Core Protein: Wo gal Endoplasmic Reticulum Golgi Apparatus ‘with Extruded i Proteoglycans is of Protein ‘Synthesis of Modification Extrusion of Proteoglycans ‘and Addition of Xyiose _—_of Extended Polysaccharide Chain ‘Through Membrane Figure 2-30 Biosynthesis pathway for mammalian proteoglycans and associated GAGs. ‘-Phosphoadenosine-: = ‘S’-phosphosulfate (PAPS) Figure 2-31 Structure of PAPS.Other Glycoconiugates 97 Sulfation reactions take place in both the endoplasmic reticulum and the Golgi complex: The donor of the sulfate group is a compound called PAPS, which is derived from ATP and sulfate (Figure 2-31), Sulfation reactions involving PAPS are also important in drug metabolism; the attachment of a sulfate improves the aqueous solubility ofa drug or its metabolites, allowing for better excretion. Other Glycoconjugates ‘As noted earlier, sugar molecules are attach roles in molecular recognition processes and stabilizing and p biomolecules. This section describes some of the most important ¢ sugars are attached. Glycolipids Glycolipids, which consist of sugars attached to lipids, of their structures are presented in Chapter 5. Sphingolipi include a subclass (glycosphingolipids) in which sugars are attached (Figure 2-32). A single sugar may be attached, or there may be longer chains, ed to a wide variety of other biomolecules, playing rerhaps protecting these other Jasses of biomolecules where are found in biomembranes. Details ids are a general class of lipids chat to a ceramide molecule some with 4 4 po A “ig “NAcylated Sphingosine (Ceramide) ? Galp(t—4) 61661-9197 ENDIF = 2 AAQYYWrnm™’”’: e carves waa nee 07 Nef SYYYYYr (3) on 2 & ‘Sia jure 2-32 Gl i ic ia Hse adi lay ates The abbreviation Sia is used to indicate a sialic acid, here38 Chapter 2. Carbohydrates branches. The combination of ¢ globoside is ceramide with a chai rather complex sugar chain attach The sugar moiety or moieties membrane, and these groups are Hexosidases are enzymes that sugar residue at a time. These enzy! Defects in these enzymes can | Table 2-2 lists several of these gh involved in each. Glycoproteins Many cellular proteins are modified th Unlike proteoglycans Bh being carbohydrate. chains; these are glycoproteins. 2 small fraction of their mass generally shorter, to the chains found in proteoglyca hormone and erythropoietin, in may also be glycosylated; for exampl rrticular asparat lypeptide ass through the side chain o rbohydrates) or to oxygens in ked carbohydrates); see Figure attachment of a sugar chain to a pa Following protein synthesis, pol endoplasmic reticulum and later p may be attached to a nitrogen in (so-called N-linked cat residues (so-called O-lin endoplasmic reticulum and in the Golgi complex, rydrate chains destined to complex. Complex carbohy using a special lipid, dolichol phosphate, embe reticulum (Figure 2-34). The assembled chain is t tachment ofa single sugar residue to the target protein, and the zed enzymes. After processing in the endoplasmic reticulum, ‘modified in the Golgi complex. Finally, these glycoproteins carbohydrate chains start with att chains are elongated by special the glycoproteins may be further ceramide and @ in of a couple .ed. Gangliosi of a glycolipid frequently involved down ce a on found in the lysosomal lead to serious ycolipid storage 4 contain more branching, ins, In mat G creted proteins lycoproxcins may ako be found 35 so fg in lactalbumin). Soluble proteins in the ce le, ribonuclease . led a fe sugar residue is cal cerebros single gd a ganglioside ceramide wie: of ug Tevalent in the membranes ofa ides fed from the surface of the roject ourwal 7 Pred in cell-cell recognicion cee and gangliosides, removin, compartments ote pidoses), such a5 Tay-Sachs dict identifies the enzyme defen” cat, pathologies ( tachment of one OF MOTE Carbohydr, Neoprotins are moslY PON, with ony wyeofne attached carbohydrate chaing a and have more diversity in sugar sequence compare nd ae pone cell-surface PFOCSINS aTe BYCOsyLeg {oa hormones such as thyroid-stimulatng rough the 2 B differs from ribonuclease A only in the gine on this enzyme. chains in eukaryotes enter the lumen of the the Golgi complex. During this passage, sugar, f asparagine residues on the polypeptide the side chains of serine or threonine 2-33. N-links are formed in the while O-links are formed only in the Golgi be attached to asparagine are first assembled .dded in the membrane of the endoplasmic hen transferred to the target protein. Simpler are sorted in the Golgi complex, to direct the proteins to their proper cellular locations—for example, the plasma membrane or internal organelles such as lysosomes. Disease Lipid Accumulated Main Organs Affected Enzyme Deficiency Fabry Ceramide trihexoside Kidney appalicalak Gaucher Glucocerebroside Brain, liver, spleen Glucosylceramide Krabbe Galactocerebroside. Brain ee a alactosylceramit Tay-Sachs Ganglioside GM, Brain B-p-galactosidase B-p-Hexosaminidase A Table 2-2 Glycolipid Storage DiseasesOther Giycoconjugates 90 Protein Chain Asparagine HOH ‘Side Chain Ho: £2 u NLinked Sugar Ho, NH—C—CH,-CH in H ond: H Hs NAcotyigucosamine Protein Chain ‘O-Linked Sugar N-Acetyigalactosamine Figure 2-33 O- and N-linked sugars in proteoglycans. Figure 2-34 Dolichol phosphate. Lectins Lecting are proteins that specifically bind carbohydrate chains on the outside of cells to make particular cell-to-cell contacts. One class of lectins, C-type lectins, requires calcium ions to help form protein-carbohydrate complexes. Bacteria use lectins to adhere to epithelial cells in the gut, binding to oligosaccharides on the cell surface. Viruses enter cells by using the protein hemagglutinin to bind to sialic acid residues on glycoproteins embedded in the cel] membrane. The “H” part of the designation of strains of influenza virus (e.g., the HINT strain of the influenza virus, commonly referred to as swine flu) refers to hemagglutinin; several different, but closely related forms of this viral protein exist. The “N” part of the nomenclature for the influenza virus refers to neuraminidase, another viral protein of which there are several variants. Neuraminidase is an enzyme that cleaves the glycosidic bond joining the sialic residue to the embedded protein after virus enters cell—an action that frees the virus for unpackaging and replication.40 Chapter2 Carbohydrates Clinical Applications Clotting and Heparan/ Heparin Heparin is usually prepared from intestinal mucosa, which is rich in heparan sulfate proteoglycans, Heparin is a fraction of heparan. Its structure is that of a repeating disaccharide of glucosamine and iduronate, heavily sulfated. A pentameric sequence within this glycosaminoglycan has Sreat affinity for antithrombin III, a plasma protein that inhibits proteases involved in forming blood clots. This pentasaccharide contains a rare 3-O-sulfated glucosamine (Figure 2-35). Complexation of heparin with antithrombin enhances the activity of antithrombin greatly, so heparin—and the pentasaccharide in particular—is used in anticoagulant therapy. Blood Group Antigens ‘The membranes of red blood cells contain glycolipids that carry oligosaccharides recognized by the immune system. The lipid is a derivative of sphingosine (see Chapters 5 and 13 for more details on glycolipids). The oligosaccharide is composed of a glucose linked directly to the lipid, followed by a galactose residue, an N-acetylglucosamine residue, and then a galactose residue. There is usually a terminal fucose residue in a 1 —> 2 linkage. The exact sequence of sugars, and the manner in which they are linked and branched by glycosyltransferase enzymes, determines the blood group type (Figure 2-36). One type of glycosyltransferase attaches a branching N-acetylgalactosamine to the galactose residue toward the end of the chain, giving the “A” blood type. Another type attaches galactose instead, giving the “B” blood type. In the “O” blood type, neither enzyme is active, and the chain is not branched. A rare blood type is produced by a deficiency in the enzyme that attaches the L-fucose; this is the “I” blood type. Other rare blood types are caused by linking the fucose in a 1 —> 4 manner rather than a 1 —> 2 linkage. The immune system can produce antibodies against these various short oligosaccharides, People with A-type blood will have antibodies against the B-type oligosaccharides; conversely, people with B-type blood will produce antibodies against the A-type antigen. The antibodies can cause blood cells to clump together, leading to serious circulatory problems. For this reason, it is essential in blood transfusions to match the type of the blood donor to that of the blood recipient. People with the O-type antigens are called universal dcnors, in recognition of the fact that people with A- or B-type blood lack antibodies to the O antigen. Note, however, that people with O-type blood must receive that type in a transfusion, and not A- or B-type blood, because the A and B antigens would be recognized and attacked by the host immune system, Figure 2-35 Heparin pentasaccharide.Clinical Applications — 44 Coramise} ~[Gie-Gar-GanacrGa Typo ‘Ceramide | Gio-Gal-GaiNthc-Gal (en) Type A Ht Fucose Figure 2-36 _Glycolipids for the A-B-O blood types and the structure of fucose, an unusual sugar in these glycolipids. Blood Glucose Levels, Hemoglobin Glycosylation, and Diabetes The reducing sugar glucose has a free aldehyde when it is in the linear (not ring) form; this aldehyde is reactive with amino groups, so glucose can be covalently and nonenzymatically linked to a protein through the side-chain amino group of a lysine residue or the terminal —NH; of the protein chain. The extent of glycosylation of a common protein, such as hemoglobin (Hb) in circulating erythrocytes, depends on the concentration of free glucose. Glucose levels in blood are unusually high in the disease diabetes mellitus, so that Hb becomes glycosylated. Glycosylated Hb is denoted as HbA,; the major sugar derivative comes from the reaction of glucose with the amino terminal group on the B subunit of Hb. This product, which is denoted as HbA,,, can be detected and quantified using suitable clinical assays. The lifetime of a typical erythrocyte is 110 to 120 days,so the amount of glycosylated Hb is a record of the average level of glucose in the blood over the previous couple of months. A high level of HbA,, indicates high blood glucose levels, signifying that blood glucose is not being well controlled; conversely, one check for satisfactory control of blood glucose in diabetes mellitus is the reduction in detected HbA,.. Action of Penicillin A major component of bacterial cell walls is peptidoglycan, which is made of short peptide chains joined to a long heteropolysaccharide of alternating N-acetylglucosamine and N-acetylmuramic acid residues. The peptide chains are unusual in that they often contain a large proportion of p-amino acids, instead of the usual L-amino acids. These short peptide chains act as cross-links between polysaccharide chains. The cross-linking reaction is catalyzed by peptidoglycan transpeptidase. It is this enzyme that is the target of the antibiotic penicillin. Without a strong, intact cell wall, the bacteria cannot survive, and penicillin weakens the cell wall by inhibiting the cross-linking reaction.42 Chapter 2 Carbohydrates CH,OH 3 HO: 8 . PetpeTone Figure 2-37 a, B-Trehalose. Arrows point to the at and B linkages: c M. Petitou, B. Casu, and U. Lindahl. (2003). Figure 2-38 Vitamin C. QUESTIONS FOR DISCUSSION 1. Is glycerol a carbohydrate? Is it a monosaccharide? What about acetone? : : Figure 2-37 gives the structure of trehalose. Predict the products of hydrolysis of this sugar, Is trehalose a reducing sugar? What about the products? 3. In solution, fructose exists about 60% in the pyranose form; of this 60%, 57% is the B-p-fructopyranose,and only about 3% is the c-p-fructopyranose. Why is the B anomer favored? 4. Suggest a structure for the product of the reaction of glucose with a primary amine (as in the N-terminal valine in the beta subunit of hemoglobin). Figure 2-38 presentsa structure for ascorbic acid, vitamin C. Is vitamin C a carbohydrate? 6. Why does reduction of v-fructose’give a mixture of p-sorbitol and p-mannitol? (See Figure 2-12.) 7. Heparin preparations have reportedly been contaminated with chondroitin sulfate (CS). Such contamination is quite serious, as it can lead to severe allergic reactions. The contaminating CS was oversulfated (co a degree not found naturally), and the presence of this contaminant was likely the result of deliberate adulteration by the supplier of the heparin. Compare the structures of heparin and CS, and comment on how CS could pass as heparin in cursory tests of the properties of the heparin preparation, v REFERENCES G. Gahmberg and M. Tolvanen. (1996). “w i glycoproteins,” Trends Biochem. 08-311.” mammation cell surface proteins are Morrison and R. N. Boyd. (1992). Organic i ike }oyd. (1992). Organic Chemistry, 6th ed., Prentice = Hall, Englewood “1976-1983, a criti - x binding shan gata! Petiod in the history of heparin: The discovery of the antithrombi fockime 85, imie 85:83-89,