Identification of the rat Heymann nephritis autoantigen (GP330) as a receptor site for plasminogen

John J. Kanalas, Sudesh P Makker

Research output: Contribution to journalArticle

84 Citations (Scopus)

Abstract

Previous results have demonstrated the binding of a 76- and 80-kDa serum protein to the Heymann nephritis autoantigen, gp330. This 76-kDa serum protein was purified by column chromatography and preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis. A rabbit polyclonal antibody for the serum protein was produced and used to screen a rat liver cDNA expression library. Sequence analysis of an isolated clone identified the serum protein as plasminogen. Plasminogen was isolated from rat serum by standard techniques, and the binding of plasminogen to gp330 was confirmed by Western analysis. Enzyme-linked immunosorbent assay results demonstrated a time-dependent, saturable, and specifically inhibitable binding of plasminogen to gp330. There was no significant difference in the binding of the two carbohydrate forms of plasminogen to gp330. Plasminogen binding to gp330 could be completely inhibited by the addition of exogenous gp330. This binding could also be partially inhibited by benzamidine but only slightly by the lysine analogue, ε-aminocaproic acid. However, even a combination of these two inhibitors could not completely block the binding of plasminogen to gp330 indicating that gp330 may be binding to plasminogen through some other unknown interactions. These results demonstrate that gp330 is a receptor site for plasminogen.

Original languageEnglish (US)
Pages (from-to)10825-10829
Number of pages5
JournalJournal of Biological Chemistry
Volume266
Issue number17
StatePublished - 1991
Externally publishedYes

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Membranous Glomerulonephritis
Plasminogen
Autoantigens
Rats
Blood Proteins
Aminocaproic Acid
Immunosorbents
Column chromatography
Electrophoresis
Gene Library
Sodium Dodecyl Sulfate
Liver
Lysine
Sequence Analysis
Chromatography
Polyacrylamide Gel Electrophoresis
Assays
Complementary DNA
Clone Cells
Enzyme-Linked Immunosorbent Assay

ASJC Scopus subject areas

  • Biochemistry

Cite this

Identification of the rat Heymann nephritis autoantigen (GP330) as a receptor site for plasminogen. / Kanalas, John J.; Makker, Sudesh P.

In: Journal of Biological Chemistry, Vol. 266, No. 17, 1991, p. 10825-10829.

Research output: Contribution to journalArticle

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