A cellulose-binding domain-fused recombinant human T cell connective tissue-activating peptide-III manifests heparanase activity

Biochem Biophys Res Commun. 1999 Feb 24;255(3):657-62. doi: 10.1006/bbrc.1999.0181.

Abstract

The chemokine connective tissue-activating peptide (CTAP)-III, which belongs to the leukocyte-derived growth factor family of mediators, was previously shown to be mitogenic for fibroblasts. However, it has recently been shown that CTAP-III, released from platelets, can act like a heparanase enzyme and degrade heparan sulfate. This suggests that CTAP-III may also function as a proinflammatory mediator. We have successfully cloned CTAP-III from a lambdagt11 cDNA library of PHA-activated human CD4(+) T cells and produced recombinant CTAP-III as a fusion protein with a cellulose-binding domain moiety. This recombinant CTAP-III exhibited heparanase activity and released degradation products from metabolically labeled, naturally produced extracellular matrix. We have also developed polyclonal and monoclonal antibodies, and these antibodies against the recombinant CTAP-III detected the CTAP-III molecule in human T cells, polymorphonuclear leukocytes, and placental extracts. Thus, our study provides tools to examine further immune cell behavior in inflamed sites rich with extracellular moieties and proinflammatory mediators.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal / immunology
  • Bacterial Proteins / genetics*
  • CD4-Positive T-Lymphocytes / enzymology
  • CD4-Positive T-Lymphocytes / metabolism*
  • Carrier Proteins / genetics*
  • Cellulose / metabolism*
  • Cloning, Molecular
  • Escherichia coli / genetics
  • Extracellular Matrix / metabolism
  • Glucuronidase*
  • Glycoside Hydrolases / metabolism*
  • Heparitin Sulfate / metabolism
  • Humans
  • Mice
  • Mice, Inbred BALB C
  • Neutrophils / enzymology
  • Neutrophils / metabolism
  • Peptides / genetics*
  • Peptides / metabolism
  • Precipitin Tests
  • Protein Binding / genetics*
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism

Substances

  • Antibodies, Monoclonal
  • Bacterial Proteins
  • Carrier Proteins
  • Peptides
  • Recombinant Fusion Proteins
  • cellulose binding protein A, Clostridium cellulovorans
  • connective tissue-activating peptide
  • Cellulose
  • Heparitin Sulfate
  • Glycoside Hydrolases
  • heparanase
  • Glucuronidase