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

Hanry Yu, Christopher V. Nicchitta, Janardan Kumar, Michael Becker, Itaru Toyoshima, Michael Sheetz

Research output: Contribution to journalArticle

51 Scopus citations

Abstract

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 α 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.

Original languageEnglish (US)
Pages (from-to)171-183
Number of pages13
JournalMolecular Biology of the Cell
Volume6
Issue number2
DOIs
StatePublished - Jan 1 1995
Externally publishedYes

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

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