Detection of O -GlcNAc modifications on cardiac myofilament proteins

Genaro A. Ramirez-Correa; Isabel Martinez Ferrando; Gerald Hart; Anne Murphy

doi:10.1007/978-1-62703-386-2_13

Detection of O -GlcNAc modifications on cardiac myofilament proteins

Genaro A. Ramirez-Correa, Isabel Martinez Ferrando, Gerald Hart, Anne Murphy

School of Medicine

Research output: Chapter in Book/Report/Conference proceeding › Chapter

1 Scopus citations

Abstract

In this chapter it is described a general method that has been used successfully by more than one laboratory interested in detecting O -GlcNAc in myofilament proteins. Alternative reagents for chemo-enzymatic or metabolic labeling will be indicated, as well as references for more details in alternative methods. The outline is divided into (1) Enrichment of O -GlcNAc Stoichiometry, (2) Cardiac Myofilament Protein Isolation, (3) SDS-PAGE, (4) "Reduction and Alkylation," (5) In-Gel Protein Digestion, (6) Chemo-enzymatic Labeling of O -GlcNAc Moieties (Click Chemistry), (7) Biotin Alkyne Tagging, (8) Strong Cation Exchange (SCX) and Streptavidin, and (9) b -Elimination and Michael Addition (BEMAD) for O -GlcNAc Site-Mapping.

Original language	English (US)
Title of host publication	Heart Proteomics
Subtitle of host publication	Methods and Protocols
Publisher	Humana Press Inc.
Pages	157-168
Number of pages	12
ISBN (Print)	9781627033855
DOIs	https://doi.org/10.1007/978-1-62703-386-2_13
State	Published - 2013

Publication series

Name	Methods in Molecular Biology
Volume	1005
ISSN (Print)	1064-3745

Keywords

Cardiac myofilaments
Chemo-enzymatic labeling
Click chemistry
Michael addition
O -GlcNAcylation
O-linked- β - D -N-acetylglucosamine
Posttranslational modification
β -Elimination

ASJC Scopus subject areas

Molecular Biology
Genetics

Access to Document

10.1007/978-1-62703-386-2_13

Cite this

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title = "Detection of O -GlcNAc modifications on cardiac myofilament proteins",

abstract = "In this chapter it is described a general method that has been used successfully by more than one laboratory interested in detecting O -GlcNAc in myofilament proteins. Alternative reagents for chemo-enzymatic or metabolic labeling will be indicated, as well as references for more details in alternative methods. The outline is divided into (1) Enrichment of O -GlcNAc Stoichiometry, (2) Cardiac Myofilament Protein Isolation, (3) SDS-PAGE, (4) {"}Reduction and Alkylation,{"} (5) In-Gel Protein Digestion, (6) Chemo-enzymatic Labeling of O -GlcNAc Moieties (Click Chemistry), (7) Biotin Alkyne Tagging, (8) Strong Cation Exchange (SCX) and Streptavidin, and (9) b -Elimination and Michael Addition (BEMAD) for O -GlcNAc Site-Mapping.",

keywords = "Cardiac myofilaments, Chemo-enzymatic labeling, Click chemistry, Michael addition, O -GlcNAcylation, O-linked- β - D -N-acetylglucosamine, Posttranslational modification, β -Elimination",

author = "Ramirez-Correa, {Genaro A.} and Ferrando, {Isabel Martinez} and Gerald Hart and Anne Murphy",

year = "2013",

doi = "10.1007/978-1-62703-386-2_13",

language = "English (US)",

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series = "Methods in Molecular Biology",

publisher = "Humana Press Inc.",

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AU - Ferrando, Isabel Martinez

AU - Hart, Gerald

AU - Murphy, Anne

PY - 2013

Y1 - 2013

N2 - In this chapter it is described a general method that has been used successfully by more than one laboratory interested in detecting O -GlcNAc in myofilament proteins. Alternative reagents for chemo-enzymatic or metabolic labeling will be indicated, as well as references for more details in alternative methods. The outline is divided into (1) Enrichment of O -GlcNAc Stoichiometry, (2) Cardiac Myofilament Protein Isolation, (3) SDS-PAGE, (4) "Reduction and Alkylation," (5) In-Gel Protein Digestion, (6) Chemo-enzymatic Labeling of O -GlcNAc Moieties (Click Chemistry), (7) Biotin Alkyne Tagging, (8) Strong Cation Exchange (SCX) and Streptavidin, and (9) b -Elimination and Michael Addition (BEMAD) for O -GlcNAc Site-Mapping.

AB - In this chapter it is described a general method that has been used successfully by more than one laboratory interested in detecting O -GlcNAc in myofilament proteins. Alternative reagents for chemo-enzymatic or metabolic labeling will be indicated, as well as references for more details in alternative methods. The outline is divided into (1) Enrichment of O -GlcNAc Stoichiometry, (2) Cardiac Myofilament Protein Isolation, (3) SDS-PAGE, (4) "Reduction and Alkylation," (5) In-Gel Protein Digestion, (6) Chemo-enzymatic Labeling of O -GlcNAc Moieties (Click Chemistry), (7) Biotin Alkyne Tagging, (8) Strong Cation Exchange (SCX) and Streptavidin, and (9) b -Elimination and Michael Addition (BEMAD) for O -GlcNAc Site-Mapping.

KW - Cardiac myofilaments

KW - Chemo-enzymatic labeling

KW - Click chemistry

KW - Michael addition

KW - O -GlcNAcylation

KW - O-linked- β - D -N-acetylglucosamine

KW - Posttranslational modification

KW - β -Elimination

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ER -

Detection of O -GlcNAc modifications on cardiac myofilament proteins

Abstract

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Keywords

ASJC Scopus subject areas

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