TY - JOUR
T1 - Human endothelial cell aminopeptidase P
AU - Ju, H.
AU - Denslow, N. D.
AU - Ryan, J. W.
AU - Papapetropoulos, A.
AU - Antonov, A.
AU - Virmani, R.
AU - Kolodgie, F. D.
AU - Gerritv, R. G.
AU - Catravas, J. D.
N1 - Copyright:
Copyright 2006 Elsevier B.V., All rights reserved.
PY - 1996
Y1 - 1996
N2 - On sequencing peptides formed by LysC digests of guinea pig lung and kidney aminopeptidase P (AmP). we found peptide segments that are highly homologous with three of six conserved blocks characteristic of the protine peptidase family (PPF). It. as we postulate, AmP is a member of the PPF, the order of the conserved blocks (block A is most N-terminal, block F is most C-terminal) provides clues for the design of oligonucleotide primers for cloning a substantial portion of AmP cDNA. Thus, we prepared two forward and two reverse primers: N - representing a novel sequence believed to be closest to the cDNA 5 -end, C- and -C representing block C, and -EF representing parts of blocks E and F (which are virtually fused in AmP). We also used the 3 RACE kit APT primer. RT-PCR was performed using human aortic endothelial cell (HAEC) RNA. NK APT primers produced a 1,000 bp product; N-, -C yielded a 320 bp product; N-, -EF yielded a 630 bp product. We conclude that HAEC contain AmP mRNA. Judging from the MW of the AmP peptide, we have obtained the cDNA that encodes its C-terminal half plus an unlranslated peptide. The N-, -EF segment contains about 200 amino acid residues; the N-, -C segment contains about 100 residues; F is followed by about 100 residues, some probably removed in translation.
AB - On sequencing peptides formed by LysC digests of guinea pig lung and kidney aminopeptidase P (AmP). we found peptide segments that are highly homologous with three of six conserved blocks characteristic of the protine peptidase family (PPF). It. as we postulate, AmP is a member of the PPF, the order of the conserved blocks (block A is most N-terminal, block F is most C-terminal) provides clues for the design of oligonucleotide primers for cloning a substantial portion of AmP cDNA. Thus, we prepared two forward and two reverse primers: N - representing a novel sequence believed to be closest to the cDNA 5 -end, C- and -C representing block C, and -EF representing parts of blocks E and F (which are virtually fused in AmP). We also used the 3 RACE kit APT primer. RT-PCR was performed using human aortic endothelial cell (HAEC) RNA. NK APT primers produced a 1,000 bp product; N-, -C yielded a 320 bp product; N-, -EF yielded a 630 bp product. We conclude that HAEC contain AmP mRNA. Judging from the MW of the AmP peptide, we have obtained the cDNA that encodes its C-terminal half plus an unlranslated peptide. The N-, -EF segment contains about 200 amino acid residues; the N-, -C segment contains about 100 residues; F is followed by about 100 residues, some probably removed in translation.
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M3 - Article
AN - SCOPUS:33749159541
SN - 0892-6638
VL - 10
SP - A625
JO - FASEB Journal
JF - FASEB Journal
IS - 3
ER -