Overexpression of a human prostate-specific glandular kallikrein, hK2, in E. coli and generation of antibodies.
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.
Saedi
M S
MS
Hybritech Incorporated, San Diego, CA 92196, USA.
Cass
M M
MM
Goel
A S
AS
Grauer
L
L
Hogen
K L
KL
Okaneya
T
T
Griffin
B Y
BY
Klee
G G
GG
Young
C Y
CY
Tindall
D J
DJ
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
D000911:Antibodies, Monoclonal; D018076:DNA, Complementary; D011993:Recombinant Fusion Proteins; D007610:Kallikreins; D020840:Tissue Kallikreins