Mechanistic Studies on Phenylalanine Hydroxylase from Chromobacterium violaceum. Evidence for the Formation of an Enzyme–Oxygen Complex

Stephen O. Pember; Kenneth A. Johnson; Joseph J. Villafranca; Stephen J. Benkovic

doi:10.1021/bi00431a024

Mechanistic Studies on Phenylalanine Hydroxylase from Chromobacterium violaceum. Evidence for the Formation of an Enzyme–Oxygen Complex

Stephen O. Pember, Kenneth A. Johnson, Joseph J. Villafranca, Stephen J. Benkovic

Chemistry

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42 Scopus citations

Abstract

Steady-state kinetic analysis of pterin-dependent phenylalanine hydroxylase from Chromobacterium violaceum indicated that the enzyme follows a partially ordered reaction mechanism. The data suggested that oxygen is the first substrate to bind to the enzyme. This result was further supported by rapid-quench experiments in which the enzyme–oxygen complex was trapped to yield product. Additional support for the presence of an enzyme–oxygen complex was derived from magnetic susceptibility measurements of molecular oxygen in the presence and absence of cuprous phenylalanine hydroxylase. The magnetic susceptibility of dissolved oxygen decreased in the presence of the enzyme, supporting a direct oxygen–metal interaction.

Original language	English (US)
Pages (from-to)	2124-2130
Number of pages	7
Journal	Biochemistry
Volume	28
Issue number	5
DOIs	https://doi.org/10.1021/bi00431a024
State	Published - Mar 1989

All Science Journal Classification (ASJC) codes

Biochemistry

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10.1021/bi00431a024

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title = "Mechanistic Studies on Phenylalanine Hydroxylase from Chromobacterium violaceum. Evidence for the Formation of an Enzyme–Oxygen Complex",

abstract = "Steady-state kinetic analysis of pterin-dependent phenylalanine hydroxylase from Chromobacterium violaceum indicated that the enzyme follows a partially ordered reaction mechanism. The data suggested that oxygen is the first substrate to bind to the enzyme. This result was further supported by rapid-quench experiments in which the enzyme–oxygen complex was trapped to yield product. Additional support for the presence of an enzyme–oxygen complex was derived from magnetic susceptibility measurements of molecular oxygen in the presence and absence of cuprous phenylalanine hydroxylase. The magnetic susceptibility of dissolved oxygen decreased in the presence of the enzyme, supporting a direct oxygen–metal interaction.",

author = "Pember, {Stephen O.} and Johnson, {Kenneth A.} and Villafranca, {Joseph J.} and Benkovic, {Stephen J.}",

year = "1989",

month = mar,

doi = "10.1021/bi00431a024",

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pages = "2124--2130",

journal = "Biochemistry",

issn = "0006-2960",

publisher = "American Chemical Society",

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T1 - Mechanistic Studies on Phenylalanine Hydroxylase from Chromobacterium violaceum. Evidence for the Formation of an Enzyme–Oxygen Complex

AU - Pember, Stephen O.

AU - Johnson, Kenneth A.

AU - Villafranca, Joseph J.

AU - Benkovic, Stephen J.

PY - 1989/3

Y1 - 1989/3

N2 - Steady-state kinetic analysis of pterin-dependent phenylalanine hydroxylase from Chromobacterium violaceum indicated that the enzyme follows a partially ordered reaction mechanism. The data suggested that oxygen is the first substrate to bind to the enzyme. This result was further supported by rapid-quench experiments in which the enzyme–oxygen complex was trapped to yield product. Additional support for the presence of an enzyme–oxygen complex was derived from magnetic susceptibility measurements of molecular oxygen in the presence and absence of cuprous phenylalanine hydroxylase. The magnetic susceptibility of dissolved oxygen decreased in the presence of the enzyme, supporting a direct oxygen–metal interaction.

AB - Steady-state kinetic analysis of pterin-dependent phenylalanine hydroxylase from Chromobacterium violaceum indicated that the enzyme follows a partially ordered reaction mechanism. The data suggested that oxygen is the first substrate to bind to the enzyme. This result was further supported by rapid-quench experiments in which the enzyme–oxygen complex was trapped to yield product. Additional support for the presence of an enzyme–oxygen complex was derived from magnetic susceptibility measurements of molecular oxygen in the presence and absence of cuprous phenylalanine hydroxylase. The magnetic susceptibility of dissolved oxygen decreased in the presence of the enzyme, supporting a direct oxygen–metal interaction.

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JO - Biochemistry

JF - Biochemistry

IS - 5

ER -

Mechanistic Studies on Phenylalanine Hydroxylase from Chromobacterium violaceum. Evidence for the Formation of an Enzyme–Oxygen Complex

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