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 journalArticle

24 Citations (Scopus)

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

Fingerprint

Tissue Kallikreins
Escherichia coli
Prostate-Specific Antigen
Antibodies
Recombinant proteins
Messenger RNA
Proteins
Kallikreins
Protein Sequence Analysis
Recombinant Proteins
Prostate
Polyacrylamide Gel Electrophoresis
Anti-Idiotypic Antibodies
Prostatic Neoplasms
Epithelium
Complementary DNA
Clone Cells
Rabbits
Carcinoma
Amino Acids

Keywords

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

ASJC Scopus subject areas

  • Biochemistry
  • Endocrinology
  • Molecular Biology
  • Endocrinology, Diabetes and Metabolism

Cite this

Saedi, M. S., Cass, M. M. J., Goel, A. S., Grauer, L., Hogen, K. L., Okaneya, T., ... Tindall, D. J. (1995). Overexpression of a human prostate-specific glandular kallikrein, hK2, in E. coli and generation of antibodies. Molecular and Cellular Endocrinology, 109(2), 237-241. https://doi.org/10.1016/0303-7207(95)03508-5

Overexpression of a human prostate-specific glandular kallikrein, hK2, in E. coli and generation of antibodies. / Saedi, Mohammad S.; Cass, Michelle M J; Goel, Amita S.; Grauer, Lana; Hogen, Kristine L.; Okaneya, Toshikazu; Griffin, Beverly Y.; Klee, George G.; Young, Charles Y F; Tindall, Donald J.

In: Molecular and Cellular Endocrinology, Vol. 109, No. 2, 01.04.1995, p. 237-241.

Research output: Contribution to journalArticle

Saedi, MS, Cass, MMJ, Goel, AS, Grauer, L, Hogen, KL, Okaneya, T, Griffin, BY, Klee, GG, Young, CYF & Tindall, DJ 1995, 'Overexpression of a human prostate-specific glandular kallikrein, hK2, in E. coli and generation of antibodies', Molecular and Cellular Endocrinology, vol. 109, no. 2, pp. 237-241. https://doi.org/10.1016/0303-7207(95)03508-5
Saedi, Mohammad S. ; Cass, Michelle M J ; Goel, Amita S. ; Grauer, Lana ; Hogen, Kristine L. ; Okaneya, Toshikazu ; Griffin, Beverly Y. ; Klee, George G. ; Young, Charles Y F ; Tindall, Donald J. / Overexpression of a human prostate-specific glandular kallikrein, hK2, in E. coli and generation of antibodies. In: Molecular and Cellular Endocrinology. 1995 ; Vol. 109, No. 2. pp. 237-241.
@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, hK2, Human kallikrein, Prostate specific antigen",
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.}",
year = "1995",
month = "4",
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",
publisher = "Elsevier Ireland Ltd",
number = "2",

}

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.

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 - hK2

KW - Human kallikrein

KW - Prostate specific antigen

UR - http://www.scopus.com/inward/record.url?scp=0028908486&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0028908486&partnerID=8YFLogxK

U2 - 10.1016/0303-7207(95)03508-5

DO - 10.1016/0303-7207(95)03508-5

M3 - Article

C2 - 7664987

AN - SCOPUS:0028908486

VL - 109

SP - 237

EP - 241

JO - Molecular and Cellular Endocrinology

JF - Molecular and Cellular Endocrinology

SN - 0303-7207

IS - 2

ER -