Intracellular

Traffic &
Sorting of
Proteins
Ribosome
s:
 Overview
• How proteins are targeted to their correct destinations? :
• Clinical conditions associated with defects in protein targeting
• Drugs
• Mechanisms of certain toxins
• Maintenance of quality control in protein traffic
• Vesicle transport
• Degradation of proteins in proteasomes
A major sorting decision is made early during protein synthesis
Secretory pathway-
mechanism
 Gunter
Blobel
• There is no difference between
structure of free and bound
ribosomes

• Selection of mRNA to the ER

membrane is not via direct
binding of the mRNA itself, but
rather via binding of its nascent
translation product

• Signal hypothesis
 Signal peptide enables the binding of ‘bound ribosome’
-The Signal Hypothesis
• N-terminal
• 13-36 residues
• 6-15 hydrophobic core flanked by hydrophilic
residues
• 1 or 2 basic residues near the N-terminal
• Small and neutral residues near the cleavage
site
Signal
hypothesis:
 Transmembrane proteins are of different
classes:

Signal
sequence
Type I Transmembrane
Protein
Type II, III &IV Transmembrane
Proteins
GPI linked
proteins
Some proteins are transported post-
translationally
Quality Control in Endoplasmic Reticulum-
ERAD
Clinical
Significance
Unfolded Protein Response and
DM
From ER to Golgi and
Further
Vesicle
transport
 Coat
proteins

Golgi>>>PM/Lysosomes ER>>>Golgi Golgi>>>ER

 Fusio
• R or v-SNARE n
 Synaptobrevin
• Q or t-SNARE
 Syntaxin
 SNAP-25 [Synaptosome
Associated Protein]
• Disassembly:
SNARE

SNAP

NSF
Fusio
n
Vesicle
transport
ER resident proteins have a KDEL
sequence
• C-terminal:
 KDEL
 KKXX
 KXKXXX
• "If found, please return
to ER"
Targeting to
lysosomes
 I-cell
• Mucolipidosis II disease
• UDP-N -acetyl glucosamine
phosphotransferase
• Cultured fibroblasts-deficient in numerous
lysosomal enzymes
• Inclusions in lysosome
• These enzymes were found to be present
in excess in tissue culture media and in
extracellular fluids
• Psychomotor and skeletal defects
Cytosolic
pathway
Protein import to
peroxisomes:
• Peroxisomal matrix
targeting
sequences
• PTS-1:
 SKL
• PTS-2:
 N-terminal (R/K)
(L/V/I)X5(H/Q)
(L/A)
 The Zellweger
• Zellweger cerebrohepatorenal syndrome spectrum:
• Neonatal adrenoleukodystrophy
• Infantile Refsum disease

• Peroxisomal biogenesis disorders

• Mutation in PEX genes
 Impaired plasmalogen synthesis
 Impaired very long chain fatty acid (VLCFA) beta oxidation
 Impaired alpha oxidation
Signal sequences for nuclear import are not
cleaved
• Nuclear Localisation Signal
– Pro-Pro-Lys-Lys-Lys-Arg-
Lys-Val
Protein targeting to
mitochondria
 Protein
• 76 residues degradation
• Highly conserved
• Isopeptide bonds

 Juvenile onset Parkinsonism

 HPV

• 26 S proteasome
 20S core subunit
 19S regulatory subunit
• Bortezomib and Carfilzomib
Summar
y