Protein targeting or protein sorting is the mechanism by which a cell transports proteins to the appropriate positions in the cell

or outside of it. Sorting targets can be the inner space of an organelle, any of several interior membranes, the cell's outer membrane, or its exterior via secretion. This delivery process is carried out based on information contained in the protein itself. Correct sorting is crucial for the cell; errors can lead to diseases

All proteins have a common beginning. Synthesis of all proteins, regardless of their final destination is is initiated on free ribosomes in the cytoplasmic compartment of the cell. Any differences in the fate of proteins is a consequence of targeting signals contained within the amino acid sequence of the protein itself. 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. The continuous stretch of amino acid residues in the chain that enables targeting are called signal peptides or targeting peptides

The presequences of the targeting peptide are often found at the Nterminal 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

In prokaryotes, targeting is usually achieved by an N-terminal signal sequence of about 20 mostly hydrophobic amino acids. Targeting in eukaryotes is necessarily more complex due to the multitude of internal compartments: nucleus mitochondria peroxisomes chloroplasts endoplasmic reticulum (ER) Golgi lysosomes secretory granules

post-translational targeting: some proteins are translated in the cytosol and later transported to their destination. This occurs for proteins that go to
nucleus mitochondria chloroplasts peroxisomes

co-translational targeting (secretory pathway): The N-terminal signal sequence of the protein is recognized by a signal recognition particle (SRP) while the protein is still being synthesized on the ribosome. . Co- translational targeting

occurs in

ER Golgi lysosomes plasma membrane secreted proteins

There are three major types of transport systems

Nuclear pores Translocation across membranes. Vesicle transport

Unusual since 2-way traffic:

in: proteins, DNA
x DNA & RNA polymerases x transcriptions factors x histones etc.

out: mRNA, tRNA, rRNA

Proteins are not transported through the nuclear membrane but rather through a complex pore called the nuclear pore:
comprised of about 100 different proteins proteins smaller than 20 kDa move by diffusion proteins larger than 20 kDa move by selective transport

(nuclear localization signal)

x cluster of 4-8 positively charged amino acids (example: PKKKRLV) x signal sequence binds to receptor on the pore called importin

Entry of large proteins into the nucleus requires: Nuclear Localization Signal (NLS) lys-lys-lys-arg-lys. inside the protein, near but not at the N-terminal end of protein. NLS region of protein binds to a cytosolic nuclear import receptor protein (NIR) NIR with attached NLS-protein then binds to the nuclear pore. A GTP driven reaction results in a change in the configuration of the pore that results in the translocation (movement) of the NIR-NLS-protein complex into the nucleus When there, the NIR proteins dissociate from the NLSprotein and are returned to the cytosol. As the NLS is an internal localization signal it is not removed by a specific peptidase as are ER, mitochondrial and chloroplast localization signals that are located at the N terminal end of the protein

Key concept: Protein containing signal sequence is recognized, bound, unfolded on entry and refolded on the organellar side of the membrane. This requires a set of special organellar chaperone proteins. The protein recognition signal is quite large. The entry site to mitochondria or chloroplasts is a site where the inner and outer membranes touch. This is referred to as a contact site.

The protein containing the signal sequence is synthesized in the cytoplasm. Signal sequence binds to a receptor in the organelle membrane Receptor - protein complex diffuses within membrane to a contact site. Protein is unfolded, moved across the membrane, and refolded. These operations are carried out by the protein transporter complex and its associated chaperone proteins. The signal sequence is the first part of the protein to enter the organelle. Once inside, the signal sequence is cleaved off by a specific peptidase

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

co-translational insertion of protein into or through ER membrane via attached ribosomes (rough ER):
signal sequence of 16-30 amino acids at N-

terminus (hydrophobic) emerging signal sequence of nascent protein on free ribosome binds to signal recognition particle (SRP) -- translation is arrested.
x SRPs consist of 6 proteins and one RNA molecule (7S RNA). x The SRP-signal sequence-mRNA-ribosome complex docks with receptor on ER membrane.

signal sequence crosses ER membrane. translation continues with polypeptide chain

being pulled into the ER lumen.

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 the Golgi where further processing and final sorting occurs:

 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 and subsequently
undergo exocytosis (A) fuse with vesicles to digest their contents

(B & C) rupture, causing autolysis (D)

Diseases Peroxisomal protein transport is defective in the following genetic diseases: Zellweger syndrome. Adrenoleukodystrophy (ALD). Refsum disease