Transmembrane protein topology mapping by the substituted cysteine accessibility method (SCAM™): Application to lipid-specific membrane protein topogenesis

Mikhail Bogdanov; Wei Zhang; Jun Xie; William Dowhan

doi:10.1016/j.ymeth.2004.11.002

Transmembrane protein topology mapping by the substituted cysteine accessibility method (SCAM™): Application to lipid-specific membrane protein topogenesis

Mikhail Bogdanov, Wei Zhang, Jun Xie, William Dowhan

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

117 Scopus citations

Abstract

We provide an overview of lipid-dependent polytopic membrane protein topogenesis, with particular emphasis on Escherichia coli strains genetically altered in their lipid composition and strategies for experimentally determining the transmembrane organization of proteins. A variety of reagents and experimental strategies are described including the use of lipid mutants and thiol-specific chemical reagents to study lipid-dependent and host-specific membrane protein topogenesis by substituted cysteine site-directed chemical labeling. Employing strains in which lipid composition can be controlled temporally during membrane protein synthesis and assembly provides a means to observe dynamic changes in protein topology as a function of membrane lipid composition.

Original language	English (US)
Pages (from-to)	148-171
Number of pages	24
Journal	Methods
Volume	36
Issue number	2 SPEC. ISS.
DOIs	https://doi.org/10.1016/j.ymeth.2004.11.002
State	Published - Jun 2005
Externally published	Yes

Keywords

Cysteine scanning
Maleimides
Membrane protein
Phosphatidylethanolamine
Phospholipid
Protein topology
Topogenesis

ASJC Scopus subject areas

Molecular Biology
General Biochemistry, Genetics and Molecular Biology

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10.1016/j.ymeth.2004.11.002

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title = "Transmembrane protein topology mapping by the substituted cysteine accessibility method (SCAM{\texttrademark}): Application to lipid-specific membrane protein topogenesis",

abstract = "We provide an overview of lipid-dependent polytopic membrane protein topogenesis, with particular emphasis on Escherichia coli strains genetically altered in their lipid composition and strategies for experimentally determining the transmembrane organization of proteins. A variety of reagents and experimental strategies are described including the use of lipid mutants and thiol-specific chemical reagents to study lipid-dependent and host-specific membrane protein topogenesis by substituted cysteine site-directed chemical labeling. Employing strains in which lipid composition can be controlled temporally during membrane protein synthesis and assembly provides a means to observe dynamic changes in protein topology as a function of membrane lipid composition.",

keywords = "Cysteine scanning, Maleimides, Membrane protein, Phosphatidylethanolamine, Phospholipid, Protein topology, Topogenesis",

author = "Mikhail Bogdanov and Wei Zhang and Jun Xie and William Dowhan",

note = "Funding Information: The work from the authors{\textquoteright} laboratory was supported by National Institutes of General Medical Science Grant GM20478 and the John S. Dunn, Sr. Research Foundation awarded to W.D. Phil Heacock assisted in drawing the figures.",

year = "2005",

month = jun,

doi = "10.1016/j.ymeth.2004.11.002",

language = "English (US)",

volume = "36",

pages = "148--171",

journal = "Methods",

issn = "1046-2023",

publisher = "Academic Press Inc.",

number = "2 SPEC. ISS.",

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TY - JOUR

T1 - Transmembrane protein topology mapping by the substituted cysteine accessibility method (SCAM™)

T2 - Application to lipid-specific membrane protein topogenesis

AU - Bogdanov, Mikhail

AU - Zhang, Wei

AU - Xie, Jun

AU - Dowhan, William

N1 - Funding Information: The work from the authors’ laboratory was supported by National Institutes of General Medical Science Grant GM20478 and the John S. Dunn, Sr. Research Foundation awarded to W.D. Phil Heacock assisted in drawing the figures.

PY - 2005/6

Y1 - 2005/6

N2 - We provide an overview of lipid-dependent polytopic membrane protein topogenesis, with particular emphasis on Escherichia coli strains genetically altered in their lipid composition and strategies for experimentally determining the transmembrane organization of proteins. A variety of reagents and experimental strategies are described including the use of lipid mutants and thiol-specific chemical reagents to study lipid-dependent and host-specific membrane protein topogenesis by substituted cysteine site-directed chemical labeling. Employing strains in which lipid composition can be controlled temporally during membrane protein synthesis and assembly provides a means to observe dynamic changes in protein topology as a function of membrane lipid composition.

AB - We provide an overview of lipid-dependent polytopic membrane protein topogenesis, with particular emphasis on Escherichia coli strains genetically altered in their lipid composition and strategies for experimentally determining the transmembrane organization of proteins. A variety of reagents and experimental strategies are described including the use of lipid mutants and thiol-specific chemical reagents to study lipid-dependent and host-specific membrane protein topogenesis by substituted cysteine site-directed chemical labeling. Employing strains in which lipid composition can be controlled temporally during membrane protein synthesis and assembly provides a means to observe dynamic changes in protein topology as a function of membrane lipid composition.

KW - Cysteine scanning

KW - Maleimides

KW - Membrane protein

KW - Phosphatidylethanolamine

KW - Phospholipid

KW - Protein topology

KW - Topogenesis

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DO - 10.1016/j.ymeth.2004.11.002

M3 - Article

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AN - SCOPUS:19444377006

SN - 1046-2023

VL - 36

SP - 148

EP - 171

JO - Methods

JF - Methods

IS - 2 SPEC. ISS.

ER -

Transmembrane protein topology mapping by the substituted cysteine accessibility method (SCAM™): Application to lipid-specific membrane protein topogenesis

Abstract

Keywords

ASJC Scopus subject areas

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