METABOLIC

HOMEOSTASIS
Regulation of Appetite, Energy Expenditure, Body Weight
Unit-IV
What is Metabolic Homeostasis ?
• When a normal animal overeats, the resulting additional fat somehow signals the brain to induce the
animal to eat less and to expend more energy. Conversely, the loss of fat stimulates increased eating
until the lost fat is replaced.

• Evidently, animals have a “lipo-stat” that can keep the amount of body fat constant to within 1% over
many years. At least a portion of the lipo-stat resides in the hypothalamus (a part of the brain that
hormonally controls numerous physiological functions), since damaging it can yield a grossly obese
animal.

• Despite this obvious set of controls in animals, there has been an explosion of obesity in many industrial
nations. It has, in fact, become a world health problem, leading to diabetes and heart disease. As a result
of numerous studies in recent years, researchers have been able to outline the mechanisms involved in
metabolic homeostasis, the balance between energy influx and energy expenditure, and to identify
some of the irregularities that lead to obesity.

• A variety of mutant strains of rodents have been generated that cause obesity. The study of these
mutants has resulted in the identification of several hormones that act in a coordinated manner to
regulate appetite
A. AMP-Dependent Protein Kinase Is the Cell’s Fuel Gauge
• All of the metabolic pathways are affected in one way or another by the need for ATP, as is indicated by
the cell’s AMP-to-ATP ratio. Several enzymes are either activated or inhibited allosterically by AMP, and
several others are phosphorylated by AMP-dependent protein kinase (AMPK), a major regulator of
metabolic homeostasis. AMPK activates metabolic breakdown pathways that generate ATP while
inhibiting biosynthetic pathways so as to conserve ATP for more vital processes.
• AMPK is an heterotrimer found in all eukaryotic organisms from yeast to humans. The subunit
contains a Ser/Thr protein kinase domain, and the subunit contains sites for allosteric activation by
AMP and inhibition by ATP.
• Like other protein kinases, AMPK’s kinase domain must be phosphorylated for activity. Binding of
AMP to the subunit causes a conformational change that exposes Thr 172 in the activation loop of the
subunit, promoting its phosphorylation and increasing its activity at least 100-fold. AMP can activate the
phosphorylated enzyme up to 5-fold more.
• There are two isoforms of the α subunit, two of the β subunit, and three of the γ subunit, giving rise to
12 possible heterotrimeric combinations, with splice variants yielding further diversity. The major
kinase that phosphorylates AMPK is named LKB1.The knockout of LKB1 in mouse liver results in the
loss of the phosphorylated form of AMPK
AMP dependent Protein Kinase
a. AMPK Activates Glycolysis in Ischemic Cardiac Muscle
• AMPK’s targets include the heart isozyme of the bifunctional enzyme PFK-2/FBPase-2, which controls the
fructose-2,6-bisphosphate (F2,6P) concentration.
• The phosphorylation of this isozyme activates the PFK-2 activity, increasing [F2,6P], which in turn activates
PFK-1, the rate-determining enzyme of glycolysis. Consequently, in ischemic (blood-starved) heart muscle
cells, which receive insufficient oxygen for oxidative phosphorylation to maintain adequate concentrations
of ATP, the resulting AMP build-up causes the cells to switch to anaerobic glycolysis for ATP production.

b. AMPK Inhibits Lipogenesis, Cholesterol Synthesis, and

Gluconeogenesis in Liver
• AMPK-mediated phosphorylation also inhibits acetyl-CoA carboxylase (ACC), which catalyzes the first
committed step of fatty acid synthesis, and hydroxy-methyl-glutaryl-CoA reductase (HMG-CoA reductase),
which catalyzes the rate-determining step in cholesterol biosynthesis.
• Activated AMPK inhibits gluconeogenesis in a more complicated way: It phosphorylates and thereby
inactivates the transcriptional coactivator TORC2 (for transducer of regulated CREB activity-2), which in
concert with the transcriptional activator CREB, would otherwise induce the transcription of the gene
encoding PEP carboxy-kinase (PEPCK), the enzyme that catalyzes the rate determining step of
gluconeogenesis. Consequently, when the rate of ATP production is inadequate, these biosynthetic
pathways are turned off, thereby conserving ATP for more vital cellular functions.
c. AMPK Promotes Fatty Acid Oxidation and Glucose Uptake but
Inhibits Glycogen Synthesis in Skeletal Muscle
• The inhibition of ACC results in a decrease in the concentration of malonyl-CoA, the starting material for fatty
acid biosynthesis. Malonyl-CoA has an additional role, however. It is an inhibitor of carnitine palmitoyl-
transferase I, which is required to transfer cytosolic palmitoyl-CoA into mitochondria for oxidation. The
decrease in malonyl-CoA concentration therefore allows more palmitoyl-CoA to be oxidized.
• AMPK also increases the recruitment of GLUT4 to muscle cell plasma membranes, as well as stimulating its
expression, thus facilitating the insulin-independent entry of glucose into these cells. In addition, AMPK inhibits
glycogen synthase (which catalyzes the rate-limiting reaction in glycogen synthesis). In fact, the subunit of
AMPK has a glycogen-binding domain that presumably recruits AMPK to the vicinity of glycogen synthase

d. AMPK Inhibits Fatty Acid Synthesis and Lipolysis in Adipocytes

• AMPK inhibits fatty acid synthesis in adipocytes by phosphorylating ACC as described above. Moreover, AMPK
phosphorylates hormone-sensitive triacylglycerol lipase in adipose tissue. This phosphorylation inhibits rather
than activates the enzyme, in part by preventing the relocation of the enzyme to the lipid droplet, the cellular
location of lipolysis.
• As a result, fewer triacylglycerol molecules are broken down so that fewer fatty acids are exported to the
bloodstream. This latter process seems paradoxical (fatty acid oxidation would help relieve an ATP deficit),
although it has been speculated that it prevents the cellular buildup of fatty acids to toxic levels. The major
effects of AMPK activation on glucose and lipid metabolism in liver, skeletal muscle, heart muscle, and adipose
tissue are diagrammed in Fig. 27-4
Figure: 27-4
B. Adiponectin Regulates AMPK Activity
• Adiponectin is a 247-residue protein hormone, secreted exclusively by adipocytes, that helps regulate
energy homeostasis and glucose and lipid metabolism by controlling AMPK activity. Its monomers
consist of an N-terminal collagen like domain and a C-terminal globular domain.
• Adiponectin occurs in the bloodstream in several forms: a low molecular weight (LMW) trimer formed
by the coiling of its collagen like domains into a triple helix as well as hexamers (MMW) and multimers
(HMW) that form disulfide cross-linked bouquets (Fig. 27-5). In addition, globular adiponectin, formed
by the cleavage of the collagen like domain to release globular monomers, occurs in lower
concentrations. The binding of adiponectin to adiponectin receptors, which occur on the surfaces of both
liver and muscle cells, acts to increase the phosphorylation and activity of AMPK. This, as we have seen ,
inhibits gluconeogenesis and stimulates fatty acid oxidation in liver and stimulates glucose uptake and
glucose and fatty acid oxidation in muscle.
• All of these effects act to increase insulin sensitivity, in part because adiponectin and insulin elicit
similar responses in tissues such as liver. Decreased adiponectin is associated with insulin resistance.
Paradoxically, the blood concentration of adiponectin, which is secreted by adipocytes, decreases with
increased amounts of adipose tissue.
• This may be because increased adipose tissue is also associated with increased production of tumor
necrosis factor- (TNF-), a cytokine that decreases both the expression and secretion of adiponectin from
adipose tissue.
Figure: 27-5
C. Leptin
• Two of the genes whose mutations cause obesity in mice are known as obese (ob) and diabetes (db; the
wild-type genes are designated OB and DB). Homozygotes for defects in either of these recessive genes,
ob/ob or db/db, are grossly obese and have nearly identical phenotypes (Fig. 27-6).
• Indeed, the way in which these phenotypes were distinguished was by surgically linking the circulation
of a mutant mouse to that of a normal (OB/OB) mouse, a phenomenon named parabiosis. ob/ob mice so
linked exhibit normalization of body weight and reduced food intake, whereas db/db mice do not do so.
• This suggests that ob/ob mice are deficient in a circulating factor that regulates appetite and metabolism,
whereas db/db mice are defective in the receptor for this circulating factor.
• The mouse OB gene encodes a 146-residue monomeric protein named leptin (Greek: leptos, thin; Fig. 27-
7) that has no apparent homology with proteins of known sequence. Leptin, which was discovered by
Jeffrey Friedman, is expressed only by adipocytes, which in doing so appear to inform the brain of how
much fat the body carries. Thus, injecting leptin into ob/ob mice causes them to eat less and to lose
weight.
• In fact, leptin-treated ob/ob mice on a restricted diet lost 50% more weight than untreated ob/ob mice on
the same diet, which suggests that reduced food intake alone is insufficient to account for leptin-induced
weight loss. Leptin appears to control energy expenditure as well.
Figure: 27-6
X-ray Structure of Leptin
• Leptin injection has no effect on db/db mice. The leptin receptor gene was identified by making a cDNA
library from mouse brain tissue that specifically bound leptin and then identifying a receptor-
expressing clone by its ability to bind leptin (gene cloning techniques).
• This gene, which has been shown to be the DB gene, encodes a protein named OB-R (for OB receptor)
that appears to have a single transmembrane segment and an extracellular domain that resembles the
receptors for certain cytokines (proteins that regulate the differentiation, proliferation, and activities of
various blood cells).
• OB-R protein, which was discovered by Louis Tartaglia, has at least six alternatively spliced forms that
appear to be expressed in a tissue-specific manner (alternative gene). In normal mice, the hypothalamus
expresses high levels of a splice variant of OB-R that has a 302-residue cytoplasmic segment.
• However, in db/db mice, this segment has an abnormal splice site that truncates it to only 34 residues,
which almost certainly renders this OB-R variant unable to transmit leptin signals. Thus, it appears that
leptin’s weight-controlling effects are mediated by signal transduction resulting from its binding to the
OB-R protein in the hypothalamus.
• Human leptin is 84% identical in sequence to that of mice. The use of a radioimmunoassay to measure
the serum levels of leptin in normal-weight and obese humans established that in both groups serum
leptin concentrations increase with their percentage of body fat as does the ob mRNA content of their
adipocytes. Moreover, after obese individuals had lost weight, their serum leptin concentrations and
adipocyte ob mRNA content declined.
• This suggests that most obese persons produce sufficient amounts of leptin but have developed “leptin
resistance.” Since leptin must cross the blood–brain barrier in order to exert its effects on the
hypothalamus, it has been suggested that this crossing is somehow saturable, thus limiting the
concentration of leptin in the brain. The high concentration of leptin in obese individuals is not without
affect, however.
• OB-R is also expressed in peripheral tissues where leptin has been shown to function as well. While not
preventing obesity, the hormone has been shown to directly stimulate the oxidation of fatty acids as
well as to inhibit the accumulation of lipids in non-adipose tissue. It does so by activating AMP-
dependent protein kinase (AMPK), which in turn phosphorylates and thereby inactivates
• acetyl-CoA carboxylase (ACC). This reduces the malonyl-CoA concentration, thereby decreasing its
inhibition of carnitine palmitoyl-transferase I, which then transports fatty acyl-CoA into the
mitochondrion for oxidation.
• A small minority of obese individuals have been found to be leptin deficient in a manner similar to
ob/ob mice. Two grossly obese children who are members of the same highly consanguineous
(descended from the same ancestors) family (they are cousins and both sets of parents are cousins) have
been shown to be homozygous for a defective OB gene. The children, at the ages of 8 and 2 years old,
respectively, weighed 86 and 29 kg and were noted to have remarkably large appetites.
• Their OB genes have a deletion of a single guanine nucleotide in codon 133, thereby causing a
frameshift mutation that, it is likely, renders the mutant leptin biologically inactive. Moreover, their
leptin serum levels were only 10% of normal. Leptin injections have relieved their symptoms.
a. Leptin resistance may be a contributing factor to obesity
• If leptin is produced in proportion to body-fat mass and leptin inhibits eating, why do people become
obese? Obese people, in most cases, have both functioning leptin receptor s and a high blood
concentration of leptin. The failure to respond to the anorexigenic effects of leptin is called leptin
resistance . What is the basis of leptin resistance?
• As for most questions in the exciting area of energy homeostasis, the answer is not well worked out, but
recent evidence suggests that a group of proteins called suppressors of cytokine signaling (SOCS) may
take part. These proteins fine-tune some hormonal systems by inhibiting receptor action. SOCS proteins
inhibit receptor signaling by a number of means .
• Consider, for example, the effect of SOCS proteins on the insulin receptor. Recall that insulin stimulates
the autophosphorylation of tyrosine residues on the insulin receptor , which in turn phosphorylates IRS-
1 , initiating the insulin-signaling pathway (Figure 27. 4A ). SOCS proteins bind to phosphorylated
tyrosine residues on receptors or other members of the signal-transduction pathway, thereby disrupting
signal flow and thus altering the cell’s biochemical activity (Figure 27.4B) . In other cases, the binding of
SOCS proteins to components of the signal-transduction pathway may also enhance proteolytic
degradation of these components by the proteasome.
• Evidence in support of a role for SOCS in leptin resistance comes from mice that have had SOCS
selectively deleted from POMC-expressing neurons. These mice display an enhanced sensitivity to
leptin and are resistant to weight gain even when fed a high-fat diet. The reason why the activity of
SOCS proteins increases, leading to leptin resistance, remains to be determine
FIGURE 27.4 Suppressors of cytokine signaling (SOCS) regulate receptor function.  (A) Insulin binding results in
phosphorylation of the receptor and subsequent phosphorylation of IRS-1. These processes initiate the insulin-
signaling pathway. (B) SOCS proteins disrupt interactions of components of the insulin-signaling pathway by binding
phosphorylated proteins and thereby inhibiting the pathway. The binding of a signal component by SOCS results in
proteasomal degradation in some cases. (IRS-1, insulin-receptor substrate 1; SOCS, suppressor of cytokine signaling.)
D. Insulin
• We have discussed the insulin signaling cascade and the role of insulin in peripheral tissues such as
muscle and adipose tissue in stimulating the uptake of glucose (Fig. 20-11) and its storage as glycogen or
fat.
• Insulin receptors also occur in the hypothalamus. Consequently, the infusion of insulin into rats with
insulin-deficient diabetes inhibits food intake, reversing the overeating behavior characteristic of the
disease. Knock-out mice have been developed with a central nervous system–specific disruption of the
insulin receptor gene.
• These mice have no alteration in brain development or survival but become obese, with increased body
fat, increased leptin levels, increased serum triacylglycerol, and the elevated plasma insulin levels
characteristic of insulin resistance.
• Evidently, insulin also plays a role in the neuronal regulation of food intake and body weight. As we
discuss in, insulin and leptin both act through receptors in the hypothalamus to decrease food intake.
Figure: 20-11
E. Ghrelin and PYY3–36
a. Ghrelin and PYY3–36 Act as Short-Term Regulators of Appetite:
Ghrelin, which was discovered by Masayasu Kojima and Kenji Kanagawa, is an appetite-stimulating
gastric peptide that is secreted by the empty stomach. This 28-residue peptide was first discovered and
named for its function as a growth hormone–releasing peptide (ghrelin is an abbreviation for growth-
hormone-release). Octanoylation of its Ser 3 is required for activity

Human ghrelin
X = Ser modified with n-octanoic acid
• Injection of ghrelin has been shown to induce adiposity (increased adipose tissue) in rodents by
stimulating an increase in food intake while reducing fat utilization. In humans in states of positive
energy balance such as obesity or high caloric intake, circulating ghrelin levels are decreased, whereas
during fasting, circulating ghrelin levels increase.
 PYY 3 − 36

• 500 Kcal of food during the next 24-hour period, whereas those receiving saline controls ate 2200 kcal
during the same period.

Human
F. Hypothalamic Integration of Hormonal Signals
a. Neurons of the Arcuate Nucleus Region of the Hypothalamus Integrate
and Transmit Hunger Signals
• About half of the length of the hypothalamus is taken up by the arcuate nucleus, a
collection of neuronal cell bodies consisting of two cell types: the NPY/AgRP cell type
and the POMC/CART cell type. These cell types are named after the neuropeptides
they secrete.
• Neuropeptide Y (NPY)

The C-terminal carboxyl is amidated

• is a potent stimulator of food intake and an inhibitor of energy expenditure, as is
Agouti related peptide (AgRP). Pro-opiomelanocortin (POMC) is post-translationally
processed in the hypothalamus to release -melanocyte stimulating hormone (-MSH).
Cocaine and amphetamine-regulated transcript (CART) and -MSH are both inhibitors
of food intake and stimulators of energy expenditure.
• The balance of the secretions from these two cell types is controlled by leptin, insulin, ghrelin, and
PYY3–36 (Fig. 27-8). Leptin and insulin signal satiety and therefore decrease appetite by diffusing
across the blood–brain barrier to the arcuate nucleus, where they stimulate POMC/CART neurons to
produce CART and -MSH, while inhibiting the production of NPY from NPY/AgRP neurons. Leptin
receptors act through the JAK–STAT signal transduction pathway.
• Ghrelin has receptors on NPY/AgRP neurons that stimulate the secretion of NPY and AgRP to increase
appetite. Interestingly, PYY3–36, a peptide that is homologous to NPY, binds specifically to NPY
receptor subtype Y2R on NPY/AgRP neurons. This subtype is an inhibitory receptor, however, so
binding of PYY3–36 causes a decrease in secretion from NPY/AgRP neurons. The integrated stimuli of
all these secretions from the arcuate nucleus control appetite

G. Control of Energy Expenditure by Adaptive Thermogenesis

• The energy content of food is utilized by an organism either in the performance of work or the
generation of heat. Excess energy is stored as glycogen or fat for future use. In well-balanced
individuals, the storage of excess fuel remains constant over many years.
• However, when energy consumed is consistently greater than energy expended, obesity results. The
body has several mechanisms for preventing obesity. One of them, as discussed above, is appetite
control. The other is diet-induced thermogenesis, a form of adaptive thermogenesis (heat production
in response to environmental stress).
• adaptive thermogenesis in response to cold, which occurs in rodents and newborn humans through
the uncoupling of oxidative phosphorylation in brown adipose tissue.
• The mechanism of this thermogenesis involves the release of norepinephrine from the brain in response
to cold, its binding to -adrenergic receptors on brown adipose tissue inducing an increase in [cAMP],
which in turn initiates an enzymatic phosphorylation cascade that activates hormone-sensitive
triacylglycerol lipase.
• The resulting increase in the concentration of free fatty acids provides fuel for oxidation as well as
inducing the opening of a proton channel, called uncoupling protein-1 (UCP1) or thermogenin, in the
inner mitochondrial membrane. The opening of UCP1 discharges the proton gradient across the inner
mitochondrial membrane, thus uncoupling electron transport from ATP production. The energy that
would otherwise have been used to drive ATP synthesis is thereby released as heat.
• Although metabolic measurements in adult humans clearly demonstrate that an increase in energy
intake causes an increase in metabolic rate and thermogenesis, the cause of this increase is unclear.
Adult humans have little brown adipose tissue. However, skeletal muscle represents up to 40% of their
total body weight and has high mitochondrial capacity.
• Homologs of UCP1 have been identified: UCP2 occurs in many tissues including white adipose tissue,
whereas UCP3 occurs in brown adipose tissue, white adipose tissue, and muscle. Leptin has been
shown to upregulate UCP2. However, it has yet to be demonstrated that UCP3 in muscle participates in
diet-induced thermogenesis. ATP-hydrolyzing substrate cycles such as that between fatty acids and
triacylglycerol in adipose tissue may also be involved.
H. Did Leptin Evolve as a Thrifty Gene?
• The unusual behavior of leptin, which serves to control weight in normal-weight individuals while its
concentration continues to climb without apparent effect in obese individuals, has led to the proposal
that leptin evolved as a “thrifty gene.”
• In hunter-gatherer societies, it was a distinct advantage to be able to survive intermittent famines. In
order to do this, fat must be stored in adipose tissue in times of plenty, making short-term obesity
advantageous. However, the accumulation of fatty acids and lipids in non-adipose tissue results in
coronary artery disease, insulin resistance, and diabetes.
• Leptin, by directly stimulating the oxidation of fatty acids as well as inhibiting the accumulation of
lipids in non-adipose tissue, is thought to protect against these diseases during short-term obesity,
thereby providing an evolutionary advantage.
• However, in recent times in industrialized nations, the unprecedented availability of food and lack of
famine has made obesity a long-term rather than a short term condition, which is a liability rather than a
benefit
REFERENCES:
• Biochemistry by Donald Voet and Judith Voet
• Biochemistry by Lubert and Stryer