Effect of fibroblast activation protein and alpha2-antiplasmin cleaving enzyme on collagen types I, III, and IV

Arch Biochem Biophys. 2007 Jan 15;457(2):177-86. doi: 10.1016/j.abb.2006.11.006. Epub 2006 Nov 17.

Abstract

The circulating enzyme, alpha2-antiplasmin cleaving enzyme (APCE), has very similar sequence homology and proteolytic specificity as fibroblast activation protein (FAP), a membrane-bound proteinase. FAP is expressed on activated fibroblasts associated with rapid tissue growth as in embryogenesis, wound healing, and epithelial-derived malignancies, but not in normal tissues. Its presence on stroma suggests that FAP functions to remodel extracellular matrix (ECM) during neoplastic growth. Precise biologic substrates have not been defined for FAP, although like APCE, it cleaves alpha2-antiplasmin to a derivative more easily cross-linked to fibrin. While FAP has been shown to cleave gelatin, evidence for cleavage of native collagen, the major ECM component, remains indistinct. We examined the potential proteolytic effects of FAP or APCE alone and in concert with selected matrix metalloproteinases (MMPs) on collagens I, III, and IV. SDS-PAGE analyses demonstrated that neither FAP nor APCE cleaves collagen I. Following collagen I cleavage by MMP-1, however, FAP or APCE digested collagen I into smaller peptides. These peptides were analogous to, yet different from, those produced by MMP-9 following MMP-1 cleavage. Amino-terminal sequencing and mass spectrometry analyses of digestion mixtures identified several peptide fragments within the sequences of the two collagen chains. The proteolytic synergy of APCE in the cleavage of collagen I and III was not observed with collagen IV. We conclude that FAP works in synchrony with other proteinases to cleave partially degraded or denatured collagen I and III as ECM is excavated, and that derivative peptides might function to regulate malignant cell growth and motility.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antigens, Neoplasm / chemistry*
  • Biomarkers, Tumor / chemistry*
  • Collagen Type I / chemistry*
  • Collagen Type II / chemistry*
  • Collagen Type IV / chemistry*
  • Gelatinases
  • Humans
  • Mass Spectrometry
  • Membrane Proteins
  • Mice
  • Molecular Sequence Data
  • Peptides / analysis
  • Rats
  • Serine Endopeptidases / chemistry*

Substances

  • Antigens, Neoplasm
  • Biomarkers, Tumor
  • Collagen Type I
  • Collagen Type II
  • Collagen Type IV
  • Membrane Proteins
  • Peptides
  • Serine Endopeptidases
  • antiplasmin-cleaving enzyme, human
  • fibroblast activation protein alpha
  • Gelatinases