title: Overexpression of a human prostate-specific glandular kallikrein, hK2, in E. coli and generation of antibodies.
abstract: The genomic and the cDNA clones of human glandular kallikrein (hK2), a member of the kallikrein family, have been isolated; however, the hK2 protein has not yet been identified and characterized. The deduced sequence of hK2 is highly homologous to prostate specific antigen (PSA), a widely accepted prognostic indicator of prostate carcinoma. Also, hK2 mRNA, like PSA mRNA, is exclusively expressed in prostatic epithelia. These two properties make hK2 a potentially useful marker for studying prostate cancer. In this paper, we describe for the first time the overexpression of the entire hK2 protein (pre-pro hK2:pphK2) in the E. coli system. Our system yields high levels of authentic pphK2 (as determined by partial amino acid sequence analysis) comprising about 40% of total cellular protein. pphK2 was purified to near homogeneity by preparative SDS/PAGE and used to generate anti-pphK2 antibodies in rabbits. The antibodies recognize the recombinant hK2 protein and a major band of approximately 34 kDa in seminal fluid.
authors.affiliation: Hybritech Incorporated, San Diego, CA 92196, USA.
authors.forename: M S
authors.initials: MS
authors.lastname: Saedi
authors.forename: M M
authors.initials: MM
authors.lastname: Cass
authors.forename: A S
authors.initials: AS
authors.lastname: Goel
authors.forename: L
authors.initials: L
authors.lastname: Grauer
authors.forename: K L
authors.initials: KL
authors.lastname: Hogen
authors.forename: T
authors.initials: T
authors.lastname: Okaneya
authors.forename: B Y
authors.initials: BY
authors.lastname: Griffin
authors.forename: G G
authors.initials: GG
authors.lastname: Klee
authors.forename: C Y
authors.initials: CY
authors.lastname: Young
authors.forename: D J
authors.initials: DJ
authors.lastname: Tindall
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000302:isolation & purification; D001483:Base Sequence; D018076:DNA, Complementary / Q000235:genetics; D004926:Escherichia coli; D005786:Gene Expression Regulation; D006801:Humans; D007610:Kallikreins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D008969:Molecular Sequence Data; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000276:immunology; D020840:Tissue Kallikreins
chemical_list: D000911:Antibodies, Monoclonal; D018076:DNA, Complementary; D011993:Recombinant Fusion Proteins; D007610:Kallikreins; D020840:Tissue Kallikreins