Characterization of kinectin, a kinesin-binding protein: primary sequence and N-terminal topogenic signal analysis.

Journal Article (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

  • PMC275827

International Standard Serial Number (ISSN)

  • 1059-1524

Digital Object Identifier (DOI)

  • 10.1091/mbc.6.2.171


  • eng

Conference Location

  • United States