Structure of the 116-kDa polypeptide of the clathrin-coated vesicle/synaptic vesicle proton pump

A 116-kDa polypeptide has recently been found to be a common component of vacuolar proton pumps isolated from a variety of sources. The 116-kDa subunit of the proton pump was purified from clathrin-coated vesicles of bovine brain, and internal sequences were obtained from proteolytic peptides. Oligo...

Full description

Saved in:
Bibliographic Details
Published in:The Journal of biological chemistry 1991-02, Vol.266 (6), p.3877-3881
Main Authors: Perin, M S, Fried, V A, Stone, D K, Xie, X S, Südhof, T C
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Aminoacids, peptides. Hormones. Neuropeptides
Analytical, structural and metabolic biochemistry
Animals
Base Sequence
Biological and medical sciences
Biological Transport
Blotting, Northern
brain
Brain Chemistry
Cattle
Clathrin - metabolism
Coated Pits, Cell-Membrane - metabolism
Fundamental and applied biological sciences. Psychology
genes
Molecular Sequence Data
Proteins
Proteins - genetics
RNA - analysis
synaptic vesicles
Vacuolar Proton-Translocating ATPases
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:A 116-kDa polypeptide has recently been found to be a common component of vacuolar proton pumps isolated from a variety of sources. The 116-kDa subunit of the proton pump was purified from clathrin-coated vesicles of bovine brain, and internal sequences were obtained from proteolytic peptides. Oligonucleotide probes designed from these peptide sequences were utilized in polymerase chain reactions to isolate partial bovine cDNA clones for the protein. Sequences from these were then utilized to isolate rat brain cDNA clones containing the full-length coding region. RNA blots indicate the presence of an abundant 3.9-kilobase message for the 116-kDa subunit in brain, and primer extension analysis demonstrates that the cloned sequence is full-length. The rat cDNA sequences predict synthesis of a protein of 96,267 Da. Analysis of the deduced amino acid sequence of the 116-kDa subunit suggests that it consists of two fundamental domains: a hydrophilic amino-terminal half that is composed of greater than 30% charged residues, and a hydrophobic carboxyl-terminal half that contains at least six transmembrane regions. The structural properties of the 116-kDa proton pump polypeptide agree well with its proposed function in coupling ATP hydrolysis by the cytoplasmic subunits to proton translocation by the intramembranous components of the pump.
ISSN:0021-9258
1083-351X
DOI:10.1016/S0021-9258(19)67875-0