The Caenorhabditis elegans unc-13 gene product is a phospholipid-dependent high-affinity phorbol ester receptor

Biochem J. 1992 Nov 1;287 ( Pt 3)(Pt 3):995-9. doi: 10.1042/bj2870995.

Abstract

The Caenorhabditis elegans unc-13 mutant is a member of a class of mutants that have un-coordinated movement. Mutations of the unc-13 gene cause diverse defects in C. elegans, including abnormal neuronal connections and modified synaptic transmission in the nervous system. unc-13 cDNA encodes a protein (UNC-13) of 1734 amino acid residues with a predicted molecular mass of 198 kDa and sequence identity to the C1/C2 regions but not to the catalytic domain of the ubiquitously expressed protein kinase C family [Maruyama & Brenner (1991) Proc. Natl. Acad. Sci. U.S.A. 88, 5729-5733]. To characterize the phorbol ester binding site of the UNC-13 protein, cDNA encoding the C1/C2-like regions (amino acid residues 546-940) was expressed in Escherichia coli and the 43 kDa recombinant protein was purified. Phorbol ester binding to the 43 kDa protein was zinc- and phospholipid-dependent, stereospecific and of high affinity (Kd 67 nM). UNC-13 specific antisera detected a protein of approx. 190 kDa in wild-type (N2) but not in mutant (e1019) C. elegans cell extracts. We conclude that UNC-13 represents a novel class of phorbol ester receptor.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Binding Sites / genetics
  • Caenorhabditis elegans / genetics*
  • Caenorhabditis elegans / metabolism
  • Caenorhabditis elegans Proteins*
  • Carrier Proteins
  • Cloning, Molecular
  • Consensus Sequence
  • Escherichia coli
  • Molecular Sequence Data
  • Peptide Fragments / genetics
  • Phorbol Esters / metabolism*
  • Phospholipids / metabolism*
  • Protein Kinase C*
  • Receptors, Drug / genetics*
  • Receptors, Drug / metabolism
  • Sequence Homology, Amino Acid
  • Zinc / metabolism

Substances

  • Caenorhabditis elegans Proteins
  • Carrier Proteins
  • Peptide Fragments
  • Phorbol Esters
  • Phospholipids
  • Receptors, Drug
  • phorbol ester binding protein
  • phorbol ester receptor
  • Protein Kinase C
  • Zinc