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. 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.
Perin
M S
MS
Howard Hughes Medical Institute, University of Texas Southwestern Medical Center, Dallas 75235.
Fried
V A
VA
Stone
D K
DK
Xie
X S
XS
Südhof
T C
TC
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001692:Biological Transport; D015152:Blotting, Northern; D001923:Brain Chemistry; D002417:Cattle; D002966:Clathrin / Q000378:metabolism*; D003034:Coated Pits, Cell-Membrane / Q000378:metabolism*; D008969:Molecular Sequence Data; D011506:Proteins / Q000235:genetics*; D012313:RNA / Q000032:analysis; D025262:Vacuolar Proton-Translocating ATPases
D002966:Clathrin; D011506:Proteins; C067470:proton pump polypeptide 116K; D012313:RNA; D025262:Vacuolar Proton-Translocating ATPases