Identification of monoclonal antibodies that recognize different disulfide bonded forms of thrombospondin 1

Lisa J. Matthias, Ian Gotis-Graham, P. Anne Underwood, H. Patrick McNeil, Philip J. Hogg*

*Corresponding author for this work

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

Thrombospondin 1 (TSP1) is a multidomain glycoprotein from platelets and cells which functions in cell-cell and cell-matrix interactions, The structure of TSP1 is regulated by sulfhydryl-disulfide interchange in the carboxy-terminal Ca2+-binding loops and globular domain which markedly influence its interaction with cell surface integrins and its inhibition of neutrophil enzymes. We have identified murine monoclonal antibodies that recognized different disulfide-bonded forms of TSP1, made by preparing TSP1 in buffers containing either 0.1 mM or 2 mM Ca2+. Antibody HB8432 recognizes TSP1 prepared in buffers containing either 0.1 or 2 mM Ca2+ while antibodies D4.6 and A65M recognized only TSP1 prepared in buffers containing 0.1 mM Ca2+. The antibodies recognize these different TSP1 preparations either adsorbed to plastic or extracellular matrix. Immunohistochemistry of human rheumatoid synovial tissue using HB8432 resulted in staining of numerous blood vessel walls and matrix cells, while D4.6 and A65M stained a subset of the HB8432 positive blood vessels and only occasionally stained matrix cells. These results suggested that different disulfide-bonded forms of TSP1 were being expressed in different areas of inflamed tissue.

Original languageEnglish
Pages (from-to)138-144
Number of pages7
JournalBiochimica et Biophysica Acta - Protein Structure and Molecular Enzymology
Volume1296
Issue number2
DOIs
Publication statusPublished - 5 Sep 1996
Externally publishedYes

Keywords

  • Cell-matrix interaction
  • Glycoprotein, multidomain
  • Human rheumatoid synovial fluid
  • Inflammation
  • Monoclonal antibody
  • Sulfhydryl-disulfide interchange
  • Thrombospondin 1

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