Endoproteolytic processing of integrin pro-alpha subunits involves the redundant function of furin and proprotein convertase (PC) 5A, but not paired basic amino acid converting enzyme (PACE) 4, PC5B or PC7

Biochem J. 2000 Feb 15;346 Pt 1(Pt 1):133-8.

Abstract

Several integrin alpha subunits undergo post-translational endoproteolytic processing at pairs of basic amino acids that is mediated by the proprotein convertase furin. Here we ask whether other convertase family members can participate in these processing events. We therefore examined the endoproteolysis rate of the integrin subunits pro-alpha5, alpha6 and alphav by recombinant furin, proprotein convertase (PC)5A, paired basic amino acid converting enzyme (PACE)4, PC1, PC2 and PC7 in vitro and/or ex vivo after overexpression in LoVo cells that were deficient in furin activity. We found that 60-fold more PC1 than furin was needed to produce 50% cleavage of pro-alpha subunit substrates in vitro; the defective pro-alpha chain endoproteolysis in LoVo cells was not rescued by overexpression of PC1 or PC2. No endoproteolysis occurred with PC7 either in vitro or ex vivo, although similar primary sequences of the cleavage site are found in integrins and in proteins efficiently processed by PC7, which suggests that a particular conformation of the cleavage site is required for optimal convertase-substrate interactions. In vitro, 50% cleavage of pro-alpha subunits was obtained with one-third of the amount of PC5A and PACE4 than of furin. In LoVo cells, PC5A remained more active than furin, PACE4 activity was quite low, and PC5B, which differs from PC5A by a C-terminal extension containing a transmembrane domain, was very inefficient in processing integrin alpha-subunit precursors. In conclusion, these results indicate that integrin alpha-subunit endoproteolytic processing involves the redundant function of furin and PC5A and to a smaller extent PACE4, but not of PC1, PC2, PC5B or PC7.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Motifs
  • Amino Acid Sequence
  • Calcium / pharmacology
  • Furin
  • Gene Expression
  • Humans
  • Hydrolysis / drug effects
  • Integrins / chemistry
  • Integrins / metabolism*
  • Kinetics
  • Membrane Proteins*
  • Molecular Weight
  • Precipitin Tests
  • Proprotein Convertase 5
  • Proprotein Convertases
  • Protein Precursors / chemistry
  • Protein Precursors / metabolism*
  • Protein Processing, Post-Translational* / drug effects
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Serine Endopeptidases / chemistry
  • Serine Endopeptidases / genetics
  • Serine Endopeptidases / metabolism*
  • Substrate Specificity
  • Subtilisins / deficiency
  • Subtilisins / genetics
  • Subtilisins / metabolism*
  • Transfection
  • Tumor Cells, Cultured

Substances

  • Integrins
  • Membrane Proteins
  • Protein Precursors
  • Recombinant Proteins
  • PCSK6 protein, human
  • Proprotein Convertase 5
  • Proprotein Convertases
  • Serine Endopeptidases
  • Subtilisins
  • proprotein convertase PC7
  • FURIN protein, human
  • Furin
  • Calcium