Identification of the rate-limiting step in serine proteinases from the effect of temperature on steady-state kinetics

Paul F. Fitzpatrick

doi:10.1016/0167-4838(89)90081-2

Identification of the rate-limiting step in serine proteinases from the effect of temperature on steady-state kinetics

Paul F. Fitzpatrick

Department of Biochemistry & Structural Biology

Research output: Contribution to journal › Article › peer-review

2 Scopus citations

Abstract

The effect of temperature on the steady-state kinetics of porcine pancreatic elastase can be used to determine whether acylation or deacylation is the rate-limiting step in catalysis. If acylation is rate-limiting, k_cat and k_cat/K_m will show the same temperature dependence. If deacylation is rate-limiting, k_cat will show a greater temperature dependence than k_cat/K_m. The temperature dependence of the steady-state kinetic parameters of t-Boc-Ala-Ala-Pro-Ala p-nitroanilide and N-acetyl-Ala-Ala-α-Aza-Ala p-nitrophenyl ester have been determined and are consistent with this prediction.

Original language	English (US)
Pages (from-to)	201-203
Number of pages	3
Journal	Biochimica et Biophysica Acta (BBA)/Protein Structure and Molecular
Volume	995
Issue number	2
DOIs	https://doi.org/10.1016/0167-4838(89)90081-2
State	Published - Apr 6 1989

Keywords

Elastase, Kinetics, Proteinase
Temperature

ASJC Scopus subject areas

Biophysics
Structural Biology
Biochemistry
Molecular Biology

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10.1016/0167-4838(89)90081-2

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title = "Identification of the rate-limiting step in serine proteinases from the effect of temperature on steady-state kinetics",

abstract = "The effect of temperature on the steady-state kinetics of porcine pancreatic elastase can be used to determine whether acylation or deacylation is the rate-limiting step in catalysis. If acylation is rate-limiting, kcat and kcat/Km will show the same temperature dependence. If deacylation is rate-limiting, kcat will show a greater temperature dependence than kcat/Km. The temperature dependence of the steady-state kinetic parameters of t-Boc-Ala-Ala-Pro-Ala p-nitroanilide and N-acetyl-Ala-Ala-α-Aza-Ala p-nitrophenyl ester have been determined and are consistent with this prediction.",

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AU - Fitzpatrick, Paul F.

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N2 - The effect of temperature on the steady-state kinetics of porcine pancreatic elastase can be used to determine whether acylation or deacylation is the rate-limiting step in catalysis. If acylation is rate-limiting, kcat and kcat/Km will show the same temperature dependence. If deacylation is rate-limiting, kcat will show a greater temperature dependence than kcat/Km. The temperature dependence of the steady-state kinetic parameters of t-Boc-Ala-Ala-Pro-Ala p-nitroanilide and N-acetyl-Ala-Ala-α-Aza-Ala p-nitrophenyl ester have been determined and are consistent with this prediction.

AB - The effect of temperature on the steady-state kinetics of porcine pancreatic elastase can be used to determine whether acylation or deacylation is the rate-limiting step in catalysis. If acylation is rate-limiting, kcat and kcat/Km will show the same temperature dependence. If deacylation is rate-limiting, kcat will show a greater temperature dependence than kcat/Km. The temperature dependence of the steady-state kinetic parameters of t-Boc-Ala-Ala-Pro-Ala p-nitroanilide and N-acetyl-Ala-Ala-α-Aza-Ala p-nitrophenyl ester have been determined and are consistent with this prediction.

KW - Elastase, Kinetics, Proteinase

KW - Temperature

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Identification of the rate-limiting step in serine proteinases from the effect of temperature on steady-state kinetics

Abstract

Keywords

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