Expression of human 60-kD heat shock protein (HSP60 or P1) in Escherichia coli and the development and characterization of corresponding monoclonal antibodies.
Human P1 protein, which is the homolog of the 60- to 65-kD heat shock "common" antigenic protein of numerous pathogenic organisms (synonyms: HSP60, GroEL homolog, or chaperonin), has been expressed to high level in Escherichia coli cells. A large number of well-characterized deletions of this protein spanning the entire sequence have been constructed and expressed. Methods to purify recombinant human HSP60 protein and its deletions from E. coli have been worked out. In addition, monoclonal antibodies to the human HSP60 protein have been raised and partially characterized. The availability of these materials should greatly aid in understanding the role of this highly conserved and immunologically important protein in autoimmune diseases and in cell structure and function.
Singh
B
B
Department of Biochemistry, McMaster University, Hamilton, Canada.
Gupta
R S
RS
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology*; D001483:Base Sequence; D015153:Blotting, Western; D003001:Cloning, Molecular; D003429:Cross Reactions; D004247:DNA; D004926:Escherichia coli / Q000235:genetics; D005455:Fluorescent Antibody Technique; D006360:Heat-Shock Proteins / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D006801:Humans; D006825:Hybridomas; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D016296:Mutagenesis; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D015183:Restriction Mapping
D000911:Antibodies, Monoclonal; D006360:Heat-Shock Proteins; D011994:Recombinant Proteins; D004247:DNA