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

doi:10.1016/0303-7207(95)03508-5

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

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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 language	English (US)
Pages (from-to)	237-241
Number of pages	5
Journal	Molecular and Cellular Endocrinology
Volume	109
Issue number	2
DOIs	https://doi.org/10.1016/0303-7207(95)03508-5
State	Published - 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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10.1016/0303-7207(95)03508-5

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@article{9bfc98f4621c4093a48d39decfe6c696,

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 -34 kDa in seminal fluid.",

keywords = "Antibodies, Cloning and expression, Human kallikrein, Prostate specific antigen, hK2",

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

note = "Funding Information: We would like to thank Dr. Hsiung for supplying the plasmid pPHS579 and Dr. Abhay Kumar, and Emily Guido for their helpful suggestions. We would also like to thank Craig Fenstermaker for his invaluable assistance in preparing reagents. Supported in part by NIH grants DK41995 (CYFY), CA15083 (DJT) and CA58225 (DJT and CYFY).",

year = "1995",

month = apr,

day = "1",

doi = "10.1016/0303-7207(95)03508-5",

language = "English (US)",

volume = "109",

pages = "237--241",

journal = "Molecular and Cellular Endocrinology",

issn = "0303-7207",

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TY - JOUR

T1 - Overexpression of a human prostate-specific glandular kallikrein, hK2, in E. coli and generation of antibodies

AU - Saedi, Mohammad S.

AU - Cass, Michelle M.J.

AU - Goel, Amita S.

AU - Grauer, Lana

AU - Hogen, Kristine L.

AU - Okaneya, Toshikazu

AU - Griffin, Beverly Y.

AU - Klee, George G.

AU - Young, Charles Y.F.

AU - Tindall, Donald J.

N1 - Funding Information: We would like to thank Dr. Hsiung for supplying the plasmid pPHS579 and Dr. Abhay Kumar, and Emily Guido for their helpful suggestions. We would also like to thank Craig Fenstermaker for his invaluable assistance in preparing reagents. Supported in part by NIH grants DK41995 (CYFY), CA15083 (DJT) and CA58225 (DJT and CYFY).

PY - 1995/4/1

Y1 - 1995/4/1

N2 - 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.

AB - 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.

KW - Antibodies

KW - Cloning and expression

KW - Human kallikrein

KW - Prostate specific antigen

KW - hK2

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SN - 0303-7207

VL - 109

SP - 237

EP - 241

JO - Molecular and Cellular Endocrinology

JF - Molecular and Cellular Endocrinology

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ER -

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

Abstract

Keywords

ASJC Scopus subject areas

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