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PMID: 7641682 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

The Pichia pastoris peroxisomal protein PAS8p is the receptor for the C-terminal tripeptide peroxisomal targeting signal.

The EMBO journal ·Vol. 14 ·No. 15 ·1995-08-01 ·Pages 3627-34

Terlecky SR, Nuttley WM, McCollum D, Sock E, Subramani S

Abstract

The peroxisomal targeting signal 1 (PTS1), consisting of a C-terminal tripeptide (SKL and variants), directs polypeptides to the peroxisome matrix in evolutionarily diverse organisms. Previous studies in the methylotrophic yeast Pichia pastoris identified a 68 kDa protein, PAS8p, as a potential component of the PTS1 import machinery. We now report several new properties of this molecule which, taken together, show that it is the peroxisomal PTS1 receptor. (i) PAS8p is localized to and tightly associated with the cytoplasmic side of the peroxisomal membrane, (ii) peroxisomes of wild-type, but not of pas8 delta (null) mutant, P.pastoris cells bind a PTS1-containing peptide (CRYHLKPLQSKL), (iii) CRYHLKPLQSKL can be cross-linked to PAS8p after binding at the peroxisome membrane and (iv) purified PAS8p binds CRYHLKPLQSKL with high affinity (nanomolar dissociation constant). In addition, the tetratricopeptide repeat (TPR) domain of PAS8p is identified as the PTS1 binding region.

MeSH Terms
Amino Acid Sequence Base Sequence Binding Sites Cross-Linking Reagents Cytoplasm Fungal Proteins/genetics,metabolism Intracellular Membranes/metabolism Kinetics Membrane Proteins/genetics,metabolism Microbodies/metabolism Molecular Sequence Data Peroxisome-Targeting Signal 1 Receptor Pichia/genetics,metabolism Receptors, Cytoplasmic and Nuclear/metabolism Sequence Deletion Succinimides
Chemicals
Cross-Linking Reagents Fungal Proteins Membrane Proteins PEX5 protein, Pichia pastoris Peroxisome-Targeting Signal 1 Receptor Receptors, Cytoplasmic and Nuclear Succinimides bis(sulfosuccinimidyl)suberate
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Terlecky S R
Department of Biology, University of California-San Diego, La Jolla 92093-0322, USA.
Nuttley W M
McCollum D
Sock E
Subramani S
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1995-08-01
Pages
3627-34
Language
English
Region
England
NLM ID
8208664
PMCID
PMC394437
Subset
IM
Grants
NIDDK NIH HHS · 1 F32 DK09061-01 · United States
NIDDK NIH HHS · DK 41737 · United States
External Links
PubMed source PMC Full Text View journal details
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