Analysis of conserved residues of the human puromycin-sensitive aminopeptidase

Michael W. Thompson; Louis B. Hersh

doi:10.1016/j.peptides.2003.07.012

Analysis of conserved residues of the human puromycin-sensitive aminopeptidase

Michael W. Thompson
, Louis B. Hersh

Producción científica: Article › revisión exhaustiva

17 Citas (Scopus)

Resumen

The puromycin-sensitive aminopeptidase (ApPS) is a zinc metallopeptidase involved in the degradation of neuropeptides. Putative catalytic residues of the enzyme, Cys146, Glu338, and Lys396 were mutated, and the resultant mutant enzymes characterized. ApPS C146S exhibited normal catalytic activity, ApPS E338A exhibited decreased substrate binding, and ApPS K396I exhibited decreases in both substrate binding and catalysis. ApPS K396I and ApPS Y394F were analyzed with respect to transition state inhibitor binding. No effect was seen with the K396I mutation, but ApPS Y394F exhibited a 3.3-fold lower affinity for RB-3014, a transition state inhibitor, indicating that Tyr394 is involved in transition state stabilization.

Idioma original	English
Páginas (desde-hasta)	1359-1365
Número de páginas	7
Publicación	Peptides
Volumen	24
N.º	9
DOI	https://doi.org/10.1016/j.peptides.2003.07.012
Estado	Published - sept 2003

Nota bibliográfica

Funding Information:
This work was supported by grants DA-05736 (M.W.T.) and DA-02243 (L.B.H.) from the National Institute on Drug Abuse. The inhibitor RB-3014 was kindly provided by Dr. Marie-Claude Fournie-Zaluski (Laboratoire de Pharmacochimie Moléculaire et Structurale, INSERM, Paris, France).

Financiación

This work was supported by grants DA-05736 (M.W.T.) and DA-02243 (L.B.H.) from the National Institute on Drug Abuse. The inhibitor RB-3014 was kindly provided by Dr. Marie-Claude Fournie-Zaluski (Laboratoire de Pharmacochimie Moléculaire et Structurale, INSERM, Paris, France).

Financiadores	Número del financiador
National Institute on Drug Abuse	R01DA002243

ASJC Scopus subject areas

Biochemistry
Physiology
Endocrinology
Cellular and Molecular Neuroscience

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10.1016/j.peptides.2003.07.012

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title = "Analysis of conserved residues of the human puromycin-sensitive aminopeptidase",

abstract = "The puromycin-sensitive aminopeptidase (ApPS) is a zinc metallopeptidase involved in the degradation of neuropeptides. Putative catalytic residues of the enzyme, Cys146, Glu338, and Lys396 were mutated, and the resultant mutant enzymes characterized. ApPS C146S exhibited normal catalytic activity, ApPS E338A exhibited decreased substrate binding, and ApPS K396I exhibited decreases in both substrate binding and catalysis. ApPS K396I and ApPS Y394F were analyzed with respect to transition state inhibitor binding. No effect was seen with the K396I mutation, but ApPS Y394F exhibited a 3.3-fold lower affinity for RB-3014, a transition state inhibitor, indicating that Tyr394 is involved in transition state stabilization.",

keywords = "Aminopeptidase, Bestatin, Catalysis, Catalytic residues, Mechanism, Mutagenesis, Puromycin, Substrate binding",

author = "Thompson, \{Michael W.\} and Hersh, \{Louis B.\}",

note = "Funding Information: This work was supported by grants DA-05736 (M.W.T.) and DA-02243 (L.B.H.) from the National Institute on Drug Abuse. The inhibitor RB-3014 was kindly provided by Dr. Marie-Claude Fournie-Zaluski (Laboratoire de Pharmacochimie Mol{\'e}culaire et Structurale, INSERM, Paris, France).",

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doi = "10.1016/j.peptides.2003.07.012",

language = "English",

volume = "24",

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AU - Thompson, Michael W.

AU - Hersh, Louis B.

N1 - Funding Information: This work was supported by grants DA-05736 (M.W.T.) and DA-02243 (L.B.H.) from the National Institute on Drug Abuse. The inhibitor RB-3014 was kindly provided by Dr. Marie-Claude Fournie-Zaluski (Laboratoire de Pharmacochimie Moléculaire et Structurale, INSERM, Paris, France).

PY - 2003/9

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N2 - The puromycin-sensitive aminopeptidase (ApPS) is a zinc metallopeptidase involved in the degradation of neuropeptides. Putative catalytic residues of the enzyme, Cys146, Glu338, and Lys396 were mutated, and the resultant mutant enzymes characterized. ApPS C146S exhibited normal catalytic activity, ApPS E338A exhibited decreased substrate binding, and ApPS K396I exhibited decreases in both substrate binding and catalysis. ApPS K396I and ApPS Y394F were analyzed with respect to transition state inhibitor binding. No effect was seen with the K396I mutation, but ApPS Y394F exhibited a 3.3-fold lower affinity for RB-3014, a transition state inhibitor, indicating that Tyr394 is involved in transition state stabilization.

AB - The puromycin-sensitive aminopeptidase (ApPS) is a zinc metallopeptidase involved in the degradation of neuropeptides. Putative catalytic residues of the enzyme, Cys146, Glu338, and Lys396 were mutated, and the resultant mutant enzymes characterized. ApPS C146S exhibited normal catalytic activity, ApPS E338A exhibited decreased substrate binding, and ApPS K396I exhibited decreases in both substrate binding and catalysis. ApPS K396I and ApPS Y394F were analyzed with respect to transition state inhibitor binding. No effect was seen with the K396I mutation, but ApPS Y394F exhibited a 3.3-fold lower affinity for RB-3014, a transition state inhibitor, indicating that Tyr394 is involved in transition state stabilization.

KW - Aminopeptidase

KW - Bestatin

KW - Catalysis

KW - Catalytic residues

KW - Mechanism

KW - Mutagenesis

KW - Puromycin

KW - Substrate binding

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UR - https://www.scopus.com/pages/publications/0346098044#tab=citedBy

U2 - 10.1016/j.peptides.2003.07.012

DO - 10.1016/j.peptides.2003.07.012

M3 - Article

C2 - 14706550

AN - SCOPUS:0346098044

SN - 0196-9781

VL - 24

SP - 1359

EP - 1365

JO - Peptides

JF - Peptides

IS - 9

ER -

Analysis of conserved residues of the human puromycin-sensitive aminopeptidase

Resumen

Nota bibliográfica

Financiación

ASJC Scopus subject areas

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