Dynamic O-GlcNAcylation of the small heat shock protein alpha B-crystallin

Biochemistry. 1996 Mar 19;35(11):3578-86. doi: 10.1021/bi951918j.


alphaB-Crystallin, originally described as a structural lens protein, is now known to be a member of the small heat shock protein family and is expressed in a number of nonlens tissues. This highly conserved 20 kDa protein aggregates with homologous proteins, including alphaA-crystallin and the small heat shock protein HSP28, to form large heteromeric complexes. Recently, Roquemore et al. (1992) have established that both phosphorylated and unphosphorylated forms of lens alphaB-crystallin are modified with O-linked N-acetylglucosamine, a dynamic posttranslational modification abundant on nuclear and cytoplasmic proteins. In this paper, we have identified the major site of O-GlcNAcylation on lens alphaB as Thr 170. We have further shown that this modification is not restricted to lens alphaB-crystallin but occurs on alphaB isolated from rat heart tissue and human astroglioma cells. Two-dimensional electrophoresis of rat heart alphaB-crystallin revealed two O-GlcNAcylated forms with mobilities corresponding to the unphosphorylated form (alphaB2) and an unidentified, slightly more acidic form. Phosphorylated alphaB-crystallin (alphaB1) was not detected in the rat heart preparation. The major O-GlcNAcylation site on alphaB-crystallins from rat heart also appears to be at Thr 170. Metabolic pulse-chase labeling studies of U373-MG astroglioma cells indicated that turnover of the carbohydrate on alphaB-crystallin is not static but proceeds many-fold more rapidly than turnover of the protein backbone itself, consistent with a regulatory role for O-GlcNAc on this small heat shock protein.

Publication types

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

MeSH terms

  • Acetylglucosamine / metabolism*
  • Amino Acid Sequence
  • Animals
  • Astrocytes / metabolism
  • Crystallins / metabolism*
  • Electrophoresis, Gel, Two-Dimensional
  • Heat-Shock Proteins / metabolism*
  • Humans
  • Lens, Crystalline / metabolism
  • Macaca mulatta
  • Molecular Sequence Data
  • Myocardium / metabolism
  • Peptide Fragments / chemistry
  • Protein Processing, Post-Translational
  • Rats
  • Rats, Sprague-Dawley
  • Threonine / chemistry
  • Tumor Cells, Cultured


  • Crystallins
  • Heat-Shock Proteins
  • Peptide Fragments
  • Threonine
  • Acetylglucosamine