PROTEIN SORTING and/or

PROTEIN TARGETING
A typical mammalian cell contains up to 10,000 different kinds of proteins; a
yeast cell, about 5000. For a cell to function properly, each of its numerous proteins must
be localized to the correct cellular membrane or aqueous compartment (e.g., the
mitochondrial matrix, chloroplast stroma, Lysosomal lumen, or cytosol). Hormone
receptor proteins, for example, must be delivered to the plasma membrane if the cell is to
recognize hormones, and specific ion-channel and transporter proteins are needed if the
cell is to import or export the corresponding ions and small molecules. Water-soluble
enzymes such as RNA and DNA polymerases must be targeted to the nucleus; still
others, such as proteolytic enzymes or catalase, must go to Lysosomes or peroxisomes,
respectively. Many proteins, such as hormones and components of the extra cellular
matrix, must be directed to the cell surface and secreted.

The process of directing each newly made polypeptide to a particular destination

referred to as protein targeting or protein sorting and is critical to the organization and
functioning of eukaryotic cells. This process occurs at several levels.

Protein targeting is necessary for proteins that are destined to work outside the
cytoplasm.
Protein targeting is more complex in eukaryote because of the presence of many
intracellular compartments.

Prokaryotic protein targeting (secretion):

 The chaperone protein SecB binds to the nascent polypeptide chain to prevent
premature folding which would make transport across the plasma membrane impossible.
SecE and SecY are transmembrane components which form a pore in the membrane
through which the still unfolded polypeptide is threaded. The translocation process is
energy-dependent (ATP) and is driven by SecA. Once the protein has passed through the
pore, the signal sequence is cleaved off by an extra cellular, membrane-bound protease.
Eukaryotic Protein Targeting:
The signals involved in protein sorting are also called sorting signals. They are
regions on the targeted protein with certain amino acid sequences. These signals interact
with specific receptors, either on the target organelle or a carrier protein. In the absence
of targeting signals, a protein will remain in the Cytoplasm. Examples include;

• Translational machinery
• Metabolic enzymes
• Cytoskeletal proteins
• Many signal transduction proteins

But targeting in eukaryotes is necessarily more complex due to the multitude of

internal compartments. There are two basic forms of targeting pathways, viz;
• Post translational targeting:

1. Nucleus
2. Mitochondria
3. Chloroplasts
4. peroxisomes

• Co-translational targeting or Secretary pathway:

1. Endoplasmic reticulum(ER)
2. Golgi Complex
3. Lysosomes
4. Plasma membrane
5. Secreted proteins
Post translational targeting:

Post translational targeting includes;

Nuclear targeting:

Nuclear targeting is unusually done by two-way traffic, i.e. transporting in

includes proteins, transcription factors histones etc. and out are mRNA, tRNA, rRNA.

Proteins are not transported through the nuclear membrane but rather through
complex pore called Nuclear pore, comprising of about 100 different proteins. Smaller
proteins move through pores by diffusion and larger move by selective transport, or
Nuclear Localization Signal. It is a cluster of positively charged amino acids, e.g.
PKKKRLV. Signal sequence binds to receptor on the pore called Importin.

Mitochondrial and Chloroplast targeting:

Mitochondrial targeting is not well understood and usually done by post-

translational targeting.

Co-translational targeting or Secretary pathway:

Lysosomal targeting:

Lysosomes are organelles that store enzymes which rapidly degrade other proteins
and nucleic acids. A famous target sequence is "KDEL" Initial targeting is via secretary
pathway. Final targeting occurs in the Golgi complex.
Endoplasmic reticulum targeting:

It is co-translational insertion of protein into or through ER membrane via

ribosome (RER).

A signal sequence of 16-30 with some hydrophobic amino acids at N-terminus of

nascent protein on free ribosomes binds to signal recognition particle (SRP), which halts
the translation. SRPs consist of 6 proteins and one 7S RNA molecule. The SRP-signal
sequence-mRNA-ribosomes complex docks with receptor of ER membrane and signal
sequence crosses the ER membrane. Now translation continues with polypeptide chain,
i.e. being pulled into the lumen of ER. While in the ER, many proteins undergo the first
stages of Glycosylation. Most proteins then migrate inside vesicles from the ER and enter
the cis face of Golgi complex where further processing and final sorting occurs.

The Golgi Complex:

The Golgi is responsible for further processing and final sorting of proteins. One
example is the formation of primary and secondary Lysosomes. Primary Lysosomes bud
from the trans face of the Golgi complex and subsequently undergo exocytosis (A), fuse
with vesicles to digest their contents (B & C), rupture, causing autolysis (D).
 Overview of Trafficking:

CONCLUSION
• Targeting of newly synthesized proteins is an integral component of protein synthesis.
• In prokaryotes, targeting is usually achieved by an N-terminal signal sequence of
about 20 mostly hydrophobic amino acids.
• In eukaryotes, targeting is more complex due to the large number of different cellular
compartments.
• Nuclear targeting is done via the nuclear pore using a nuclear localization signal.
• ER targeting is done via N-terminal signal sequences using SRPs with subsequent
attachment to the ER.
• Targeting is followed by transport to the Golgi complex.
• Protein degradation of damaged or obsolete proteins is carried out by Lysosomes,
vesicles filled with degradating enzymes in an ubiqutin-dependent process by specific
proteases in a large cytosolic complex called the proteasome.