Protein targeting and sorting

Protein targeting and sorting Headlines

Overview of sorting of nuclear-encoded proteins in

eukaryotic cells Two basic forms of targeting pathways Eukaryotic targeting signals to endoplasmic reticulum ( via to all secretory pathway) to mitochondria to chloroplast to peroxiosme Common principles in protein translocation across membranes

Overview of sorting of nuclear-encoded proteins in eukaryotic cells

Protein sorting

Molecular Cell Biology. Lodish et als. Figure 17-1

Question
How the cytosolic made proteins are guided towards the correct location since multiplex proteins are targeted to different organelles?

Protein traffic

Protein Trafficking/Targeting
Protein targeting is
necessary for proteins that are destined to work outside the cytoplasm.

Protein targeting is more

complex in eukaryotes because of the presence of many intracellular compartments.

Eukaryotic protein targeting

The signals involved 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 translational machinery metabolic enzymes cytoskeletal proteins many signal transduction proteins

Two basic forms of targeting pathways

post-translational targeting: nucleus mitochondria chloroplasts Peroxisomes
co-translational targeting (secretory pathway): ER Golgi lysosomes plasma membrane secreted proteins

Targeting signals

(Fig12-8 Alberts)

Eukaryotic targeting signals

Mitochondria

(Prof. Ruth Bellairs, Department of Anatomy and Developmental Biology, University College, London WC1E 6BT)

Eukaryotic targeting signals

Mitochondrial targeting signals

Eukaryotic targeting signals

Chloroplast targeting signals

1st Structure : - contain hydroxylated aa. - small hydrophobic aa. - positively charged aa. - few acidic aa.

2nd Structure: Structure - random coiled in solution - -sheet

The ability to perform conformational changes might be an important feature for cytosolic targeting

Chloroplast targeting signals. Sequence logos

(O. Emanuelsson et als. 1999 Protein Sci 1999. 8(5):978-984)

Sequence from targeting signal

Sequence of mature protein

Cleavage site for peptidase

Transport of peroxisomal proteins

How to get a folded protein across the membrane?

Eukaryotic targeting signals

The PTS1 motif

The PTS1 motif

Peroxisomal/Non-peroxisomal occurrence
AHL AKA AKF AKI AKL AKM AKV ANL ARF ARL ARM ARY ASL CKL HRL HRM KKL NKL 3/ 5 1 /19 1/ 3 1/ 9 19 / 21 3/ 1 1 /19 2/ 5 1/ 2 4/ 9 5/ 1/ 1 1 / 23 2/ 4 1/ 1 2/ 1 2/ 6 4 / 10 PHL PKL PRL SHL SKL SKI SKL SKM SKV SNL SQL SRL SRM THL TKL TKV YRM 1 /8 3/ 4 1/ 3 10 / 18 2/ 7 6/ 2 42 / 20 4/ 1 1 / 24 2/ 9 4/ 4 14 / 11 4/ 1 / 18 1 / 23 1/ 4 1 / 13

Peroxisomal membrane protein import

Types of targeting signal sequences

Target O rganelle U sual Signal Location w ithin Protein Signal R em oval* N ature of Signal

Endoplasm ic reticulum M itochondrion C hloroplast

N -term inal

(+)

C ore of 612 m ostly hydrophobic am ino acids, often preceded by one or m ore basic am ino acids 3 5 nonconsecutive Arg or Lys residues, often w ith Ser and Thr; no G lu or Asp residues N o com m on sequence m otifs; generally rich in Ser, Thr, and sm all hydrophobic am ino acid residues and poor in G lu and Asp residues U sually Ser-Lys-Leu at extrem e C -term inus O ne cluster of 5 basic am ino acids, or tw o sm aller clusters of basic residues separated by ? 10 am ino acids

N -term inal N -term inal

(+) (+)

Peroxisom e N ucleus

C -term inal Internal

(? ) (? )

Molecular Cell Biology. Lodish et als. Table17-1

Translocation across membranes

Common principles
1. Multiple pathways of protein translocation across membrane exit. Molecular chaperones are required in the cytosol, inside the organelle and often within the organelle membrane. ATP and/or GTP hydrolysis is often required. A proton-motive force across the membrane is often required. Protein translocation occurs through gated, aqueous channels

2.

3. 4. 5.