Cloning and functional expression of the cytoplasmic form of rat aminopeptidase P

G. Czirják, William A. Burkhart, Mary B. Moyer, József Antal, Stephen B. Shears, P. Enyedi

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

A rat cytoplasmic aminopeptidase P was purified from liver cytosol with a procedure including an affinity elution step with 3 μM inositol 1,3,4- trisphosphate. Proteolytic fragments were generated, sequenced and the enzyme was cloned from a rat liver cDNA library. The structure shows high (87.8% and 95.5%, respectively) sequence identity at the nucleotide and amino acid levels with the previously described human putative cytoplasmic aminopeptidase P. The cloned rat enzyme was functionally expressed in Escherichia coli and also in COS-1 cells. Western blot analysis, using an antibody generated against the recombinant protein, and Northern blot hybridization showed ubiquitous expression of the protein in different tissues with the highest expression level in the testis.

Original languageEnglish
Pages (from-to)326-336
Number of pages11
JournalBBA - Gene Structure and Expression
Volume1444
Issue number3
DOIs
Publication statusPublished - Mar 19 1999

Fingerprint

Cloning
Rats
Organism Cloning
Liver
COS Cells
Enzymes
Gene Library
Recombinant Proteins
Northern Blotting
Cytosol
Escherichia coli
Testis
Nucleotides
Western Blotting
Tissue
Amino Acids
Antibodies
X-Pro aminopeptidase
Proteins

Keywords

  • Aminopeptidase P
  • cDNA cloning
  • Cytoplasmic
  • Functional expression

ASJC Scopus subject areas

  • Biochemistry
  • Genetics
  • Structural Biology
  • Biophysics

Cite this

Cloning and functional expression of the cytoplasmic form of rat aminopeptidase P. / Czirják, G.; Burkhart, William A.; Moyer, Mary B.; Antal, József; Shears, Stephen B.; Enyedi, P.

In: BBA - Gene Structure and Expression, Vol. 1444, No. 3, 19.03.1999, p. 326-336.

Research output: Contribution to journalArticle

Czirják, G. ; Burkhart, William A. ; Moyer, Mary B. ; Antal, József ; Shears, Stephen B. ; Enyedi, P. / Cloning and functional expression of the cytoplasmic form of rat aminopeptidase P. In: BBA - Gene Structure and Expression. 1999 ; Vol. 1444, No. 3. pp. 326-336.
@article{63d18aed6a3249458727797049a5665a,
title = "Cloning and functional expression of the cytoplasmic form of rat aminopeptidase P",
abstract = "A rat cytoplasmic aminopeptidase P was purified from liver cytosol with a procedure including an affinity elution step with 3 μM inositol 1,3,4- trisphosphate. Proteolytic fragments were generated, sequenced and the enzyme was cloned from a rat liver cDNA library. The structure shows high (87.8{\%} and 95.5{\%}, respectively) sequence identity at the nucleotide and amino acid levels with the previously described human putative cytoplasmic aminopeptidase P. The cloned rat enzyme was functionally expressed in Escherichia coli and also in COS-1 cells. Western blot analysis, using an antibody generated against the recombinant protein, and Northern blot hybridization showed ubiquitous expression of the protein in different tissues with the highest expression level in the testis.",
keywords = "Aminopeptidase P, cDNA cloning, Cytoplasmic, Functional expression",
author = "G. Czirj{\'a}k and Burkhart, {William A.} and Moyer, {Mary B.} and J{\'o}zsef Antal and Shears, {Stephen B.} and P. Enyedi",
year = "1999",
month = "3",
day = "19",
doi = "10.1016/S0167-4781(99)00005-6",
language = "English",
volume = "1444",
pages = "326--336",
journal = "Biochimica et Biophysica Acta - Gene Structure and Expression",
issn = "0167-4781",
publisher = "Elsevier BV",
number = "3",

}

TY - JOUR

T1 - Cloning and functional expression of the cytoplasmic form of rat aminopeptidase P

AU - Czirják, G.

AU - Burkhart, William A.

AU - Moyer, Mary B.

AU - Antal, József

AU - Shears, Stephen B.

AU - Enyedi, P.

PY - 1999/3/19

Y1 - 1999/3/19

N2 - A rat cytoplasmic aminopeptidase P was purified from liver cytosol with a procedure including an affinity elution step with 3 μM inositol 1,3,4- trisphosphate. Proteolytic fragments were generated, sequenced and the enzyme was cloned from a rat liver cDNA library. The structure shows high (87.8% and 95.5%, respectively) sequence identity at the nucleotide and amino acid levels with the previously described human putative cytoplasmic aminopeptidase P. The cloned rat enzyme was functionally expressed in Escherichia coli and also in COS-1 cells. Western blot analysis, using an antibody generated against the recombinant protein, and Northern blot hybridization showed ubiquitous expression of the protein in different tissues with the highest expression level in the testis.

AB - A rat cytoplasmic aminopeptidase P was purified from liver cytosol with a procedure including an affinity elution step with 3 μM inositol 1,3,4- trisphosphate. Proteolytic fragments were generated, sequenced and the enzyme was cloned from a rat liver cDNA library. The structure shows high (87.8% and 95.5%, respectively) sequence identity at the nucleotide and amino acid levels with the previously described human putative cytoplasmic aminopeptidase P. The cloned rat enzyme was functionally expressed in Escherichia coli and also in COS-1 cells. Western blot analysis, using an antibody generated against the recombinant protein, and Northern blot hybridization showed ubiquitous expression of the protein in different tissues with the highest expression level in the testis.

KW - Aminopeptidase P

KW - cDNA cloning

KW - Cytoplasmic

KW - Functional expression

UR - http://www.scopus.com/inward/record.url?scp=0033583092&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033583092&partnerID=8YFLogxK

U2 - 10.1016/S0167-4781(99)00005-6

DO - 10.1016/S0167-4781(99)00005-6

M3 - Article

C2 - 10095056

AN - SCOPUS:0033583092

VL - 1444

SP - 326

EP - 336

JO - Biochimica et Biophysica Acta - Gene Structure and Expression

JF - Biochimica et Biophysica Acta - Gene Structure and Expression

SN - 0167-4781

IS - 3

ER -