Characterization of kinectin, a kinesin-binding protein: primary sequence and N-terminal topogenic signal analysis.
Published
Journal Article
Kinectin is a kinesin-binding protein (Toyoshima et al., 1992) that is required for kinesin-based motility (Kumar et al., 1995). A kinectin cDNA clone containing a 4.7-kilobase insert was isolated from an embryonic chick brain cDNA library by immunoscreening with a panel of monoclonal antibodies. The cDNA contained an open reading frame of 1364 amino acids encoding a protein of 156 kDa. A bacterially expressed product of the full length cDNA bound purified kinesin. Transient expression in CV-1 cells gave an endoplasmic reticulum distribution that depended upon the N-terminal domain. Analysis of the predicted amino acid sequence indicated a highly hydrophobic near N-terminal stretch of 28 amino acids and a large portion (326-1248) of predicted alpha helical coiled coils. The 30-kDa fragment containing the N-terminal hydrophobic region was produced by cell-free in vitro translation and found to assemble with canine pancreas rough microsomes. Cleavage of the N terminus was not observed confirming its role as a potential transmembrane domain. Thus, the kinectin cDNA encodes a cytoplasmic-oriented integral membrane protein that binds kinesin and is likely to be a coiled-coil dimer.
Full Text
Duke Authors
Cited Authors
- Yu, H; Nicchitta, CV; Kumar, J; Becker, M; Toyoshima, I; Sheetz, MP
Published Date
- February 1995
Published In
Volume / Issue
- 6 / 2
Start / End Page
- 171 - 183
PubMed ID
- 7787244
Pubmed Central ID
- 7787244
International Standard Serial Number (ISSN)
- 1059-1524
Digital Object Identifier (DOI)
- 10.1091/mbc.6.2.171
Language
- eng
Conference Location
- United States