Cloning and functional expression of the cytoplasmic form of rat aminopeptidase P.
A rat cytoplasmic aminopeptidase P was purified from liver cytosol with a procedure including an affinity elution step with 3 microM 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.
Czirják
G
G
Department of Physiology, Semmelweis University of Medicine, P.O. Box 259, H-1444, Budapest, Hungary.
Burkhart
W A
WA
Moyer
M B
MB
Antal
J
J
Shears
S B
SB
Enyedi
P
P
D000595:Amino Acid Sequence; D000626:Aminopeptidases / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000235:genetics*; D000818:Animals; D001483:Base Sequence; D019556:COS Cells; D003001:Cloning, Molecular; D003593:Cytoplasm / Q000201:enzymology; D004926:Escherichia coli / Q000235:genetics; D008099:Liver / Q000201:enzymology*; D008969:Molecular Sequence Data; D010833:Phytic Acid / Q000494:pharmacology; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats
D012333:RNA, Messenger; D010833:Phytic Acid; D000626:Aminopeptidases; C022750:X-Pro aminopeptidase