Effector proteins from P450cam and methane monooxygenase: Lessons in tuning nature's powerful reagents

Brian J. Brazeau; Bradley J. Wallar; John D. Lipscomb

doi:10.1016/j.bbrc.2003.09.242

Effector proteins from P450_cam and methane monooxygenase: Lessons in tuning nature's powerful reagents

Brian J. Brazeau, Bradley J. Wallar, John D. Lipscomb

Chemical and Structural Biology (TMED)

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

31 Scopus citations

Abstract

Effector proteins alter the kinetic or catalytic course of many oxygenase reactions. One of the first oxygenase effectors to be described was putidaredoxin, which serves to gate electron transfer into oxy-P450 _cam. In the nonheme, methane monooxygenase (MMO) system, the B-component (MMOB) serves a distinct effector function by gating substrate and oxygen into the active site of the hydroxylase component (MMOH). Here the binding parameters and binding surfaces of the MMOB-MMOH complex are determined by site-specific labeling, fluorescence titrations, chemical cross-linking, and MALDI-TOF peptide identification. Based on these data, a model for the bimolecular complex is described and a hypothesis for the structural basis for the effector function is elaborated. The bearing on the putidaredoxin effector function is discussed.

Original language	English (US)
Pages (from-to)	143-148
Number of pages	6
Journal	Biochemical and Biophysical Research Communications
Volume	312
Issue number	1
DOIs	https://doi.org/10.1016/j.bbrc.2003.09.242
State	Published - Dec 5 2003

Bibliographical note

Funding Information:
This work was supported by National Institutes of Health Grant GM40466. We thank Dr. LeeAnn Higgins and Thomas P. Krick for assistance in collecting and interpreting the mass spectrometry data and Dr. David D. Thomas for the use of the ISS K2 Fluorometer.

Keywords

Component docking
Cross-linking
Effector
Fluorescence
Kinetics
Methane monooxygenase
P450
Regulation

Publisher link

10.1016/j.bbrc.2003.09.242

Find It

Find It at U of M Libraries

Cite this

@article{99534dd0a28c42f58596c4a82d576e6b,

title = "Effector proteins from P450cam and methane monooxygenase: Lessons in tuning nature's powerful reagents",

abstract = "Effector proteins alter the kinetic or catalytic course of many oxygenase reactions. One of the first oxygenase effectors to be described was putidaredoxin, which serves to gate electron transfer into oxy-P450 cam. In the nonheme, methane monooxygenase (MMO) system, the B-component (MMOB) serves a distinct effector function by gating substrate and oxygen into the active site of the hydroxylase component (MMOH). Here the binding parameters and binding surfaces of the MMOB-MMOH complex are determined by site-specific labeling, fluorescence titrations, chemical cross-linking, and MALDI-TOF peptide identification. Based on these data, a model for the bimolecular complex is described and a hypothesis for the structural basis for the effector function is elaborated. The bearing on the putidaredoxin effector function is discussed.",

keywords = "Component docking, Cross-linking, Effector, Fluorescence, Kinetics, Methane monooxygenase, P450, Regulation",

author = "Brazeau, {Brian J.} and Wallar, {Bradley J.} and Lipscomb, {John D.}",

note = "Funding Information: This work was supported by National Institutes of Health Grant GM40466. We thank Dr. LeeAnn Higgins and Thomas P. Krick for assistance in collecting and interpreting the mass spectrometry data and Dr. David D. Thomas for the use of the ISS K2 Fluorometer. ",

year = "2003",

month = dec,

day = "5",

doi = "10.1016/j.bbrc.2003.09.242",

language = "English (US)",

volume = "312",

pages = "143--148",

journal = "Biochemical and Biophysical Research Communications",

issn = "0006-291X",

publisher = "Academic Press Inc.",

number = "1",

}

TY - JOUR

T1 - Effector proteins from P450cam and methane monooxygenase

T2 - Lessons in tuning nature's powerful reagents

AU - Brazeau, Brian J.

AU - Wallar, Bradley J.

AU - Lipscomb, John D.

N1 - Funding Information: This work was supported by National Institutes of Health Grant GM40466. We thank Dr. LeeAnn Higgins and Thomas P. Krick for assistance in collecting and interpreting the mass spectrometry data and Dr. David D. Thomas for the use of the ISS K2 Fluorometer.

PY - 2003/12/5

Y1 - 2003/12/5

N2 - Effector proteins alter the kinetic or catalytic course of many oxygenase reactions. One of the first oxygenase effectors to be described was putidaredoxin, which serves to gate electron transfer into oxy-P450 cam. In the nonheme, methane monooxygenase (MMO) system, the B-component (MMOB) serves a distinct effector function by gating substrate and oxygen into the active site of the hydroxylase component (MMOH). Here the binding parameters and binding surfaces of the MMOB-MMOH complex are determined by site-specific labeling, fluorescence titrations, chemical cross-linking, and MALDI-TOF peptide identification. Based on these data, a model for the bimolecular complex is described and a hypothesis for the structural basis for the effector function is elaborated. The bearing on the putidaredoxin effector function is discussed.

AB - Effector proteins alter the kinetic or catalytic course of many oxygenase reactions. One of the first oxygenase effectors to be described was putidaredoxin, which serves to gate electron transfer into oxy-P450 cam. In the nonheme, methane monooxygenase (MMO) system, the B-component (MMOB) serves a distinct effector function by gating substrate and oxygen into the active site of the hydroxylase component (MMOH). Here the binding parameters and binding surfaces of the MMOB-MMOH complex are determined by site-specific labeling, fluorescence titrations, chemical cross-linking, and MALDI-TOF peptide identification. Based on these data, a model for the bimolecular complex is described and a hypothesis for the structural basis for the effector function is elaborated. The bearing on the putidaredoxin effector function is discussed.

KW - Component docking

KW - Cross-linking

KW - Effector

KW - Fluorescence

KW - Kinetics

KW - Methane monooxygenase

KW - P450

KW - Regulation

UR - http://www.scopus.com/inward/record.url?scp=0345257916&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0345257916&partnerID=8YFLogxK

U2 - 10.1016/j.bbrc.2003.09.242

DO - 10.1016/j.bbrc.2003.09.242

M3 - Article

C2 - 14630032

AN - SCOPUS:0345257916

SN - 0006-291X

VL - 312

SP - 143

EP - 148

JO - Biochemical and Biophysical Research Communications

JF - Biochemical and Biophysical Research Communications

IS - 1

ER -