Ultrastructural and biochemical properties of the 120-kDa form of chick kinectin

Janardan Kumar, Harold P. Erickson, Michael Sheetz

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Kinectin, an integral membrane protein (160 kDa), was identified as a kinesin-binding protein. Analysis of the predicted amino acid sequence of kinectin cDNA indicated an α-helical coiled-coil structure from amino acid 320 to 1310. A 120-kDa kinectin has been observed consistently, and N- terminal sequencing showed that 232 amino acids were missing from the N terminus of full-length kinectin. 120-kDa kinectin was distributed in the supernatant and a low density fraction of vesicles, whereas both forms were in the high density fraction of vesicles. In the electron microscope, the 120-kDa form appeared as a linear molecule of 133 nm in length. In hydrodynamic studies, the cytosolic 120-kDa kinectin was a dimer. Monoclonal antibody molecules (anti-kinectin KR160.9) bound asymmetrically to kinectin often with two antibodies/kinectin, indicative of a parallel coiled-coil. Metabolic labeling with [3H]myristic acid showed that both the 120- and 160- kDa kinectin are myristoylated in chick embryo fibroblasts. The myristoylation of 120-kDa kinectin may provide a mechanism for linking it to a low density fraction of vesicles. Immunoprecipitation with a 160-kDa kinectin-specific antibody brought down the 120-kDa kinectin. Thus, we suggest that kinectin is an extended parallel coiled-coil dimer, often a heterodimer.

Original languageEnglish (US)
Pages (from-to)31738-31743
Number of pages6
JournalJournal of Biological Chemistry
Volume273
Issue number48
DOIs
StatePublished - Nov 27 1999
Externally publishedYes

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Amino Acids
Kinesin
Dimers
Antibodies
Protein Sequence Analysis
Myristic Acid
Chick Embryo
Hydrodynamics
Immunoprecipitation
Carrier Proteins
Membrane Proteins
Molecules
Complementary DNA
Fibroblasts
Monoclonal Antibodies
Electrons
Labeling
Electron microscopes

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Ultrastructural and biochemical properties of the 120-kDa form of chick kinectin. / Kumar, Janardan; Erickson, Harold P.; Sheetz, Michael.

In: Journal of Biological Chemistry, Vol. 273, No. 48, 27.11.1999, p. 31738-31743.

Research output: Contribution to journalArticle

Kumar, Janardan ; Erickson, Harold P. ; Sheetz, Michael. / Ultrastructural and biochemical properties of the 120-kDa form of chick kinectin. In: Journal of Biological Chemistry. 1999 ; Vol. 273, No. 48. pp. 31738-31743.
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