Identification of the phosphorylation sites of clathrin light chain LCb

J Biol Chem. 1988 Apr 25;263(12):5499-501.

Abstract

Clathrin light chains, LCa and LCb, are products of two closely related genes whose mRNAs undergo differential splicing to result in at least four different light chain isoforms. The physiological significance of clathrin light chain diversity remains unclear. To date, the only evidence for a functional distinction of LCa and LCb is the preferential phosphorylation of LCb, which takes place at serine residues and is mediated by coated vesicle-associated casein kinase II. As a first step toward determining the function of light chain diversity, we have mapped the in vitro phosphorylation sites on LCb. We use [32P]ATP to phosphorylate LCb within coated vesicles, followed by sequencing of 32P-labeled chymotryptic peptides thereof, to identify serine residues at positions 11 and 13 as the phosphorylation sites. We find that phosphorylation of LCb within coated vesicles can be inhibited by four monoclonal antibodies specific for different epitopes of the clathrin light chains.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adenosine Triphosphate / metabolism
  • Amino Acid Sequence
  • Animals
  • Brain / ultrastructure
  • Cattle
  • Chromatography, High Pressure Liquid
  • Chymotrypsin / metabolism
  • Clathrin / metabolism*
  • Coated Pits, Cell-Membrane / metabolism
  • Cysteine Endopeptidases / metabolism
  • Molecular Sequence Data
  • Peptide Fragments / metabolism
  • Phosphorylation
  • Phosphoserine / metabolism

Substances

  • Clathrin
  • Peptide Fragments
  • Phosphoserine
  • Adenosine Triphosphate
  • Chymotrypsin
  • Cysteine Endopeptidases
  • clostripain