Obtaining and characterization of EF-hand mutants of recoverin

FEBS Lett. 1998 Nov 27;440(1-2):116-8. doi: 10.1016/s0014-5793(98)01426-4.

Abstract

Several EF-hand recoverin mutants were obtained and their abilities to bind to photoreceptor membranes and to inhibit rhodopsin kinase were determined. The mutants with the 'spoiled' 2nd, 3rd or (2nd+3rd) EF-hand structures did not act upon the kinase activity in the microM range of Ca2+ concentrations. Mutations of the 4th EF hand, which 'repaired' its Ca2+-binding activity, resulted in recoverin with three 'working' Ca2+-binding sites. The latter mutant inhibited rhodopsin kinase even more effectively than the wild-type recoverin, containing two working Ca2+-binding structures.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Binding Sites
  • Calcium / metabolism
  • Calcium-Binding Proteins / chemistry
  • Calcium-Binding Proteins / genetics*
  • Calcium-Binding Proteins / metabolism*
  • Cattle
  • Eye Proteins*
  • G-Protein-Coupled Receptor Kinase 1
  • Hippocalcin
  • Lipoproteins*
  • Mutagenesis, Site-Directed
  • Mutation*
  • Nerve Tissue Proteins*
  • Phenotype
  • Phosphorylation
  • Protein Kinase Inhibitors*
  • Protein Kinases*
  • Protein Structure, Secondary
  • Recoverin
  • Retina
  • Rod Cell Outer Segment / metabolism*

Substances

  • Calcium-Binding Proteins
  • Eye Proteins
  • Lipoproteins
  • Nerve Tissue Proteins
  • Protein Kinase Inhibitors
  • Recoverin
  • Hippocalcin
  • Protein Kinases
  • G-Protein-Coupled Receptor Kinase 1
  • Calcium