Overexpression of a human prostate-specific glandular kallikrein, hK2, in E. coli and generation of antibodies

Mohammad S. Saedi, Michelle M.J. Cass, Amita S. Goel, Lana Grauer, Kristine L. Hogen, Toshikazu Okaneya, Beverly Y. Griffin, George G. Klee, Charles Y.F. Young, Donald J. Tindall

Research output: Contribution to journalArticlepeer-review

24 Scopus citations

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 -34 kDa in seminal fluid.

Original languageEnglish (US)
Pages (from-to)237-241
Number of pages5
JournalMolecular and Cellular Endocrinology
Volume109
Issue number2
DOIs
StatePublished - Apr 1 1995

Keywords

  • Antibodies
  • Cloning and expression
  • Human kallikrein
  • Prostate specific antigen
  • hK2

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Endocrinology

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