Guinea Pig Membrane-Bound Aminopeptidase P Is a Member of the Proline Peptidase Family

Denslow, Nancy D.; Ryan, James W.; Nguyen, Hamilton

doi:10.1006/bbrc.1994.2877

Cited by 15 publications

(5 citation statements)

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“…There are six putative N-linked glycosylation sites in the AP-P sequence, which are all located in the Nterminal half of the protein. Pig kidney AP-P has sequence similarities with E. coli and Streptomyces li idans AP-Ps [33,34] (44.8 % and 50.3 % similarity respectively using the GAP program [35]), especially in the C-terminal half of the protein, as well as with other members of the proline peptidase family [17,18], as previously noted [19].…”

Section: Figure 1 Nucleotide and Deduced Amino Acid Sequence Of Pig Ksupporting

confidence: 53%

“…AP-P appears to be structurally related to the ' proline peptidase ' family of hydrolytic enzymes, which includes Escherichia coli AP-P, prolidase, methionine aminopeptidase and creatinase [17,18]. The amino acid sequence of pig kidney AP-P has recently been determined by Edman degradation and MS of the purified protein [19], and sequence comparisons further emphasize the similarity between AP-P and other members of the proline peptidase family.…”

Section: Introductionmentioning

confidence: 99%

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Molecular cloning and expression in COS-1 cells of pig kidney aminopeptidase P

Hyde

Hooper

Turner

1996

Biochemical Journal

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Aminopeptidase P (AP-P; X-Pro aminopeptidase; EC 3.4.11.9), a key enzyme in the metabolism of the vasodilator bradykinin, has been cloned from a pig kidney cortex cDNA library following the use of the PCR to identify sub-libraries enriched in AP-P clones. The complete primary sequence of the enzyme has been deduced from a full-length cDNA clone. This predicts a protein of 673 amino acids with a cleavable N-terminal signal sequence and six potential N-linked glycosylation sites. A stretch of mainly hydrophobic amino acids at the C-terminus is predicted to co-ordinate the attachment of a glycosyl-phosphatidylinositol (GPI) membrane anchor. Although AP-P is a zinc metallopeptidase, the predicted primary sequence does not contain any recognizable zinc-binding motif. Transient expression of AP-P cDNA in COS-1 cells resulted in enzymic activity characteristic of AP-P, namely apstatin- and EDTA-sensitive hydrolysis of bradykinin and Gly-Pro-Hyp. The expressed protein was recognized as a polypeptide of M(r)91,000 under reducing conditions, following immunoblotting of COS-1 membranes with a polyclonal antibody raised against purified pig kidney AP-P. The presence of a GPI anchor on expressed AP-P was established by demonstrating release of the enzyme from a membrane fraction following treatment with bacterial phosphatidylinositol-specific phospholipase C and its corresponding conversion from an amphipathic to a hydrophilic form, as assessed by phase separation in Triton X-114. Sequence comparisons confirm that AP-P is a member of the proline peptidase family of hydrolytic enzymes and is unrelated in sequence to other brush-border membrane peptidases.

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Section: Figure 1 Nucleotide and Deduced Amino Acid Sequence Of Pig Ksupporting

confidence: 53%

Section: Introductionmentioning

confidence: 99%

Molecular cloning and expression in COS-1 cells of pig kidney aminopeptidase P

Hyde

Hooper

Turner

1996

Biochemical Journal

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“…Different purification methods have been described for both cytosolic APP [27,28] and mAPP. In this way, bovine [15,29], porcine [30][31][32], rat [33] and guinea pig [34,35] mAPP have been obtained from the lungs and/or kidneys, as well as from guinea pig serum [34,36]. These methods usually include several chromatographic steps, detergent use and phospholipase digestion.…”

Section: Discussionmentioning

confidence: 99%

Human recombinant membrane-bound aminopeptidase P: production of a soluble form and characterization using novel, internally quenched fluorescent substrates

Molinaro

Carmona

Juliano

et al. 2005

Biochemical Journal

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APP (aminopeptidase P) has the unique ability to cleave the N-terminal amino acid residue from peptides exhibiting a proline at P(1)'. Despite its putative involvement in the processing of bioactive peptides, among them the kinins, little is known about the physiological roles of both human forms of APP. The purpose of the present study is first to engineer and characterize a secreted form of hmAPP (human membrane-bound APP). Our biochemical analysis has shown that the expressed glycosylated protein is fully functional, and exhibits enzymic parameters similar to those described previously for mAPP purified from porcine or bovine lungs or expressed from a porcine clone. This soluble form of hmAPP cross-reacts with a polyclonal antiserum raised against a 469-amino-acid hmAPP fragment produced in Escherichia coli. Secondly, we synthesized three internally quenched fluorescent peptide substrates that exhibit a similar affinity for the enzyme than its natural substrates, the kinins, and a higher affinity compared with the tripeptide Arg-Pro-Pro used until now for the quantification of APP in biological samples. These new substrates represent a helpful analytical tool for rapid and reliable screening of patients susceptible to adverse reactions associated with angiotensin-converting enzyme inhibitors or novel vasopeptidase (mixed angiotensin-converting enzyme/neprilysin) inhibitors.

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“…3). These sequences match with the conserved sequence blocks characteristic for the proline peptidase family (Denslow et al, 1994). X-prolyl aminopeptidase has been iden tified as a zinc metalloprotease, but does not contain any of the known zinc binding sites that classify the zinc metalloproteases in distinct families (Hooper, 1994).…”

Section: Resultsmentioning

confidence: 99%

Isolation and sequence analysis of a human cDNA clone (XPNPEPL) homologous to X-prolyl aminopeptidase (aminopeptidase P)

Vanhoof

Goossens²,

Juliano³

et al. 1997

Cytogenet Genome Res

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A novel human cDNA (XPNPEPL) encoding a protein of 623 amino acids exhibiting 44% sequence identity and 62 % sequence similarity to pig kidney X-prolyl aminopeptidase (aminopeptidase P; EC 3.4.11.9) was obtained by reverse transcription/polymerase chain reaction of phytohemagglutin-in-stimulated lymphocyte mRNA. Conserved sequences were found with the prokaryotic X-prolyl aminopeptidase encoding gene (pepP). The human gene translation product exhibits a high sequence homology to the Schizosaccharomyces pombe chromosome I hypothetical protein C22G7.01c and to the S. cerevisiae ORF yll029w. Northern blot analysis indicates an ubiquitous expression of the human XPNPEPL sequence.

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Guinea Pig Membrane-Bound Aminopeptidase P Is a Member of the Proline Peptidase Family

Cited by 15 publications

References 0 publications

Molecular cloning and expression in COS-1 cells of pig kidney aminopeptidase P

Molecular cloning and expression in COS-1 cells of pig kidney aminopeptidase P

Human recombinant membrane-bound aminopeptidase P: production of a soluble form and characterization using novel, internally quenched fluorescent substrates

Isolation and sequence analysis of a human cDNA clone (XPNPEPL) homologous to X-prolyl aminopeptidase (aminopeptidase P)

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