Open AccessImmunochemical properties of a 60 kDa cell surface‐associated heat shock‐protein (Hsp60) from Helicobacter pylori
Author(s) -
Amini HamidReza,
Ascencio Felipe,
CruzVillacorta Ariel,
RuizBustos Eduardo,
Wadström Torkel
Publication year - 1996
Publication title -
fems immunology & medical microbiology
Language(s) - English
Resource type - Journals
eISSN - 1574-695X
pISSN - 0928-8244
DOI - 10.1111/j.1574-695x.1996.tb00133.x
Subject(s) - affinity chromatography , polyclonal antibodies , microbiology and biotechnology , biology , heat shock protein , monoclonal antibody , hspa14 , antibody , protein a , binding protein , hsp60 , immunoglobulin g , protein g , western blot , biochemistry , sepharose , hsp70 , enzyme , immunology , gene
Western blot analysis (immunoblotting) of cell surface‐associated proteins from Helicobacter pylori confirmed our previous findings that binding of human IgG is a common property (among H. pylori strains). Purification of the IgG‐binding proteins (IGBP) was achieved by two purification steps, affinity chromatography on IgG‐Sepharose and nickel chelate affinity chromatography. SDS‐PAGE and immunoblotting analysis revealed a 60 kDa protein with affinity for peroxidase labeled human IgG. Solid phase binding assays showed that IgG binds to an immobilized protein (IGBP). The 60 kDa IGBP binds human IgG 1 , IgG 3 and IgM. Binding could be inhibited by the kappa chain of the human IgG, but not with its Fc fragment, nor with IgA or IgM. In addition, rabbit polyclonal antibodies raised against the 60 kDa IGBP blocked IgG binding. Monoclonal antibodies, specific to the Hsp60 heat shock protein of H. pylori recognized the 60 kDa IGBP as revealed by immunoblotting analysis, both in crude preparations and in the purified fractions.