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 Citations (Scopus)

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

Fingerprint

Kinesin
Carrier Proteins
Complementary DNA
Amino Acids
Protein Sequence Analysis
Microsomes
Gene Library
Endoplasmic Reticulum
Membrane Potentials
Open Reading Frames
Canidae
Pancreas
Membrane Proteins
Clone Cells
Monoclonal Antibodies
Brain
Proteins

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

Cite this

Characterization of kinectin, a kinesin-binding protein : Primary sequence and N-terminal topogenic signal analysis. / Yu, Hanry; Nicchitta, Christopher V.; Kumar, Janardan; Becker, Michael; Toyoshima, Itaru; Sheetz, Michael.

In: Molecular Biology of the Cell, Vol. 6, No. 2, 01.01.1995, p. 171-183.

Research output: Contribution to journalArticle

Yu, Hanry ; Nicchitta, Christopher V. ; Kumar, Janardan ; Becker, Michael ; Toyoshima, Itaru ; Sheetz, Michael. / Characterization of kinectin, a kinesin-binding protein : Primary sequence and N-terminal topogenic signal analysis. In: Molecular Biology of the Cell. 1995 ; Vol. 6, No. 2. pp. 171-183.
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