PROTEIN TRAFFICKING

AND TARGETTING
• BASICS
• PROTEIN TARGETTING IN MITOCHONDRIA
• PROTEIN TARGETTING IN CHLOROPLAST
• Protein targeting refers to the methods cells use to get proteins to the
proper location after synthesis.
• Proteins play a major role in most cellular processes but must be
located properly to serve their functions.
• Knowing how newly synthesized proteins target within cells is essential
for understanding protein function.
• Proteins are synthesized either in the cytosol or on the endoplasmic
reticulum .
• When synthesized in the cytosol on free ribosomes , most proteins
diffuse freely until they are bound to a particular substrate or
assemble into a larger complex.
• Protein diffusion in the cytosol is usually rapid, so an unbound protein
is capable of diffusing across the cell in only a few seconds.
• One way cytosolic proteins are targeted within cells is by forming
large macromolecular assemblies.
TARGETTING SIGNALS
• Targeting signals are the pieces of information that enable the cellular
transport machinery to correctly position a protein inside or outside
the cell.
• There are two types of targeting peptides, the presequences and the
internal targeting peptides.
• The presequences of the targeting peptide are often found at the N-
terminal extension and is composed of between 6-136 basic and
hydrophobic amino acids.
• In case of peroxisomes the targeting sequence is on the C-terminal
extension mostly.
• Other signals, known as signal patches, are composed of
parts which are separate in the primary sequence.
• They become functional when folding brings them
together on the protein surface.
• In addition, protein modifications like glycosylations can
induce targeting.
• Conformational changes in a protein often lead to changes
in the protein's affinity toward a particular substrate.
• This process can play a crucial role in regulating the
intracellular localization of a protein.
• An example of this type of regulation is
protein phosphorylation , or addition of a phosphate
group.
• proteins targeted to mitochondria contain a specific
peptide sequence of 20–80 amino acids that mediates
their import.
• In order to allocate the many proteins at the right time to the
right place, the cell needs an address and distribution station: The
Golgi apparatus.
• There the proteins are marked and sent to the different areas of
the cell
• To reach the Golgi complex, plasma membrane, endosomes, and
lysosomes, however, proteins must enter the secretory pathway and
use membrane trafficking pathways.
Two models of protein trafficking through the
Golgi
• (A) The cisternal maturation model of protein movement through
the Golgi. As a new cis cisterna is formed it traverses the Golgi
stack, changing as it matures by accumulating medial, then trans
enzymes through vesicles that move from later to earlier cisternae
(retrograde traffic).
• (B) The vesicular transport model, where each cisterna remains in one
place with unchanging enzymes, and the proteins move forward
through the stack via vesicles that move from earlier to later cisternae
(anterograde traffic).
TWO BASIC TARGETTING PATHWAY
 COTRANSLATIONAL TRANSLOCATION
• Signal sequences are bound by a signal recognition particle as they emerge
from the ribosome. SRPs consist of 6 polypeptide and a small cytoplasmic RNA
(SRP RNA)
• The SRP binds to the ribosome and the signal sequence, inhibiting further
translation. The entire complex then binds to SRP receptors in the rough ER.
• The SRP is then released (coupled with GTP hydrolysis), and the ribosomes
binds to a membrane channel or translocon. The signal sequence is inserted
into the translocon and translation resumes
• The signal sequence is cleaved by an enzyme called signal peptidase
• Continued translation drives translocation of growing peptide across the
membrane.
• Completed polypeptides are then released into the ER lumen
POST TRANSLATIONAL
• Even though most secretory proteins are co-translationally
translocated, some are translated in the cytosol and later transported
to the ER/plasma membrane by a post-translational system.
• In prokaryotes this requires certain cofactors such as SecA and SecB.
This pathway is poorly understood in eukaryotes, but is facilitated
by Sec62 and Sec63, two membrane-bound proteins.
• In addition, proteins targeted to other destinations, such
as mitochondria, chloroplasts, or peroxisomes, use specialized post-
translational pathways.
• Also, proteins targeted for the nucleus are translocated post-
translation. They pass through the nuclear envelope via nuclear pores.