Protein sorting
BMS4002 Cell Biology
Dr Martin Figgitt
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AIMS
• Outline the processes involves protein manufacture
• Concept of protein post translational modifications
• How proteins are packages and secreted
• Concept of protein sorting
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How are proteins made?
Flow of genetic
information from
DNA to RNA to
Protein
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Central Dogma of Gene Expression
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5
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What happens to the protein
next?
• A complex series of modifications take place
• Internal protein transportation- How protein go to where are
needed?
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Key experiments by Palade & colleaguesThe Secretory
Pathway
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Conclusions from Palade’s work
These experiments provided the first evidence that
secreted proteins were synthesised into the rough ER
and moved to the Golgi and then to secretory vesicles
near the plasma membrane
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Protein sorting in eukaryotes
Proteins belong in different organelles of the cell – and
some even have their function outside the cell
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Sorting signals
• Signal sequences
• 15-60 aa
• Removed by signal peptidase when reaching the
target
• Signal patch
• Usually non-continuous stretch of sequences
• Exposed when appropriately folded
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Sorting signals
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Signal sequences
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Signal sequences
• Function: specify the direction for destination
for initial transfer to the ER: with a signal sequence at N-
terminus; consisting of 5-10 hydrophobic amino acids
• Go forward Golgi: most proteins
• Return to ER (ER residents): with a specific sequence of
4 aa at C-terminus
• Go to mitochondria: positively charged amino acids
alternate with hydrophobic ones
• Go to peroxisome: with a signal peptide of 3
characteristic at C terminus
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Key Discovery
Günter Blobel was in 1999 awarded The Nobel Prize
in Physiology or Medicine for the discovery that
"proteins have intrinsic signals that govern their
transport and localization in the cell"
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Cisternal
Fig. 5.14 (TEArt) space
Signal
sequence
Polypeptide
Ribosome
Membrane of
mRNA endoplasmic reticulum
Cytoplasm
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Fig. 5.16d (TEArt)
Protein
Vesicle Migrating Fusion
budding transport of vesicle
from rough vesicle with Golgi
endoplasmic apparatus
reticulum
Ribosome
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Fig. 5.15a (TEArt)
Secretory
vesicles
trans face
cis face
Transport vesicles
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Protein modification in the Golgi
apparatus
• Proteins and lipids manufactured on the rough and
smooth ER membranes are modified as they pass
through it
• The most common alteration is the addition or
alteration of short sugar chains, forming
glycoproteins and glycolipids
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Nucleus Plasma Extracellular fluid
Nuclear pore Protein expelled
Fig. 5.16 (TEArt) membrane
Rough
endoplasmic
reticulum
Secretory
vesicle
Ribosome
Cisternae
cis face
Proteins
trans face
Smooth Transport Golgi
endoplasmic vesicle apparatus
reticulum Cytoplasm
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Cytoplasm Endoplasmic
Fig.Phagocytosis
5.17 (TEArt) reticulum
Food Golgi
vesicle apparatus
Lysosomes Transport
vesicle
Old or damaged
Plasma Digestion of organelle
membrane phagocytized
food particles Breakdown
or cells of old
Extracellular organelle
fluid
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Transport from the Golgi Apparatus
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summary
• After protein transcription and translation
• Proteins under go a series of modifications
• There are specific signal regions that govern where the protein is to
be located
• Cell organelles eg Golgi apparatus and ER are key to protein
transportation and where proteins are to be found
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Further reading
• Read section on protein sorting and transport in any cell biology
book
• E.g. Biology by Raven.
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