Inhibition of glycosaminoglycan modification of perlecan domain I by site-directed mutagenesis changes protease sensitivity and laminin-1 binding activity

FEBS Lett. 1998 Sep 18;435(2-3):169-72. doi: 10.1016/s0014-5793(98)01063-1.

Abstract

Glycosaminoglycan attachment to perlecan domain I (173 residues) was completely prevented by site-directed mutagenesis of Ser-65, Ser-71 and Ser-76 as shown by recombinant production in mammalian cells. This did not interfere with the proper folding of the domain's SEA module but enhanced its sensitivity to neutral proteases. Lack of substitution also abolished binding to the two major heparin binding sites of laminin-1.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Binding Sites / genetics
  • Endopeptidases / metabolism*
  • Glycosaminoglycans
  • Heparan Sulfate Proteoglycans*
  • Heparitin Sulfate / analysis
  • Heparitin Sulfate / genetics*
  • Heparitin Sulfate / metabolism*
  • Laminin / metabolism*
  • Molecular Sequence Data
  • Mutagenesis, Site-Directed
  • Protein Binding / genetics
  • Proteoglycans / analysis
  • Proteoglycans / genetics*
  • Proteoglycans / metabolism*
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Substrate Specificity / genetics

Substances

  • Glycosaminoglycans
  • Heparan Sulfate Proteoglycans
  • Laminin
  • Proteoglycans
  • Recombinant Proteins
  • perlecan
  • Heparitin Sulfate
  • Endopeptidases