Interaction and biophysical properties of human lens Q155* betaB2-crystallin mutant

Mol Vis. 2005 Apr 30:11:321-7.

Abstract

Purpose: Missense mutations in crystallin genes have been identified in autosomal dominant congenital cataracts. A truncation in the CRYBB2 gene (Q155*) has been associated with cerulean cataract, however its effects on biophysical properties have not been reported. We sought to determine the changes in conformation and protein-protein interactions brought about by this mutation.

Methods: Site specific mutations were performed to obtain the Q155* betaB2-crystallin mutant. Protein-protein interactions were screened by a mammalian two-hybrid system assay. Conformational changes were studied with spectroscopy (circular dichroism and fluorescence) and FPLC chromatography.

Results: We detected a decrease in protein-protein interactions for the Q155* betaB2-crystallin mutant. The Q155* mutant shows decreased ordered structure and stability but the partially unfolded protein retains some dimer structure.

Conclusions: The Q155* mutation in betaB2-crystallin causes changes in biophysical properties that might contribute to cataract formation.

Publication types

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

MeSH terms

  • Amino Acid Sequence
  • Blotting, Western
  • Chromatography, High Pressure Liquid
  • Circular Dichroism
  • Electrophoresis, Polyacrylamide Gel
  • HeLa Cells
  • Humans
  • Molecular Sequence Data
  • Mutagenesis, Site-Directed
  • Mutation, Missense
  • Protein Binding*
  • Protein Conformation*
  • Sequence Analysis, Protein
  • Spectrometry, Fluorescence
  • Transfection
  • Two-Hybrid System Techniques
  • beta-Crystallin B Chain / chemistry*
  • beta-Crystallin B Chain / genetics

Substances

  • beta-Crystallin B Chain
  • beta-crystallin B2