Towards the N-terminal acetylome: an N-terminal acetylated peptide enrichment method using CNBr-activated sepharose resin

Xumin Zhang, Peter Højrup

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

Protein N-terminal acetylation (N(α)-acetylation) is observed widely from prokaryotes to eukaryotes. It gains increased importance in biological field, due to its multiple roles in many aspects of the protein life, such as assembly, stability, activity, and location. Today, mass spectrometry (MS) has demonstrated its unprecedented ability in a variety of proteome-wide studies including the N(α)-acetylome. The N(α)-acetylated peptides are usually immerged into the massive amount of regular peptides and are further suppressed due to their reduced positive charge states during analysis; therefore, an efficient exploration of the N(α)-acetylome necessitates a specific enrichment method. Here we describe a protocol for N(α)-acetylated peptide enrichment using CNBr-activated sepharose resin, which has proved to be simple, sensitive, and highly reproducible.
Original languageEnglish
JournalMethods in Molecular Biology
Volume981
Pages (from-to)47-56
ISSN1064-3745
DOIs
Publication statusPublished - 2013

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Sepharose
Peptides
Proteome
Acetylation
Proteins

Keywords

  • Acetylation
  • Chromatography, Liquid
  • Cyanogen Bromide
  • HeLa Cells
  • Humans
  • Peptides
  • Proteome
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Sepharose
  • Tandem Mass Spectrometry

Cite this

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title = "Towards the N-terminal acetylome: an N-terminal acetylated peptide enrichment method using CNBr-activated sepharose resin",
abstract = "Protein N-terminal acetylation (N(α)-acetylation) is observed widely from prokaryotes to eukaryotes. It gains increased importance in biological field, due to its multiple roles in many aspects of the protein life, such as assembly, stability, activity, and location. Today, mass spectrometry (MS) has demonstrated its unprecedented ability in a variety of proteome-wide studies including the N(α)-acetylome. The N(α)-acetylated peptides are usually immerged into the massive amount of regular peptides and are further suppressed due to their reduced positive charge states during analysis; therefore, an efficient exploration of the N(α)-acetylome necessitates a specific enrichment method. Here we describe a protocol for N(α)-acetylated peptide enrichment using CNBr-activated sepharose resin, which has proved to be simple, sensitive, and highly reproducible.",
keywords = "Acetylation, Chromatography, Liquid, Cyanogen Bromide, HeLa Cells, Humans, Peptides, Proteome, Reproducibility of Results, Sensitivity and Specificity, Sepharose, Tandem Mass Spectrometry",
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Towards the N-terminal acetylome : an N-terminal acetylated peptide enrichment method using CNBr-activated sepharose resin. / Zhang, Xumin; Højrup, Peter.

In: Methods in Molecular Biology, Vol. 981, 2013, p. 47-56.

Research output: Contribution to journalJournal articleResearchpeer-review

TY - JOUR

T1 - Towards the N-terminal acetylome

T2 - an N-terminal acetylated peptide enrichment method using CNBr-activated sepharose resin

AU - Zhang, Xumin

AU - Højrup, Peter

PY - 2013

Y1 - 2013

N2 - Protein N-terminal acetylation (N(α)-acetylation) is observed widely from prokaryotes to eukaryotes. It gains increased importance in biological field, due to its multiple roles in many aspects of the protein life, such as assembly, stability, activity, and location. Today, mass spectrometry (MS) has demonstrated its unprecedented ability in a variety of proteome-wide studies including the N(α)-acetylome. The N(α)-acetylated peptides are usually immerged into the massive amount of regular peptides and are further suppressed due to their reduced positive charge states during analysis; therefore, an efficient exploration of the N(α)-acetylome necessitates a specific enrichment method. Here we describe a protocol for N(α)-acetylated peptide enrichment using CNBr-activated sepharose resin, which has proved to be simple, sensitive, and highly reproducible.

AB - Protein N-terminal acetylation (N(α)-acetylation) is observed widely from prokaryotes to eukaryotes. It gains increased importance in biological field, due to its multiple roles in many aspects of the protein life, such as assembly, stability, activity, and location. Today, mass spectrometry (MS) has demonstrated its unprecedented ability in a variety of proteome-wide studies including the N(α)-acetylome. The N(α)-acetylated peptides are usually immerged into the massive amount of regular peptides and are further suppressed due to their reduced positive charge states during analysis; therefore, an efficient exploration of the N(α)-acetylome necessitates a specific enrichment method. Here we describe a protocol for N(α)-acetylated peptide enrichment using CNBr-activated sepharose resin, which has proved to be simple, sensitive, and highly reproducible.

KW - Acetylation

KW - Chromatography, Liquid

KW - Cyanogen Bromide

KW - HeLa Cells

KW - Humans

KW - Peptides

KW - Proteome

KW - Reproducibility of Results

KW - Sensitivity and Specificity

KW - Sepharose

KW - Tandem Mass Spectrometry

U2 - 10.1007/978-1-62703-305-3_5

DO - 10.1007/978-1-62703-305-3_5

M3 - Journal article

C2 - 23381853

VL - 981

SP - 47

EP - 56

JO - Methods in Molecular Biology

JF - Methods in Molecular Biology

SN - 1064-3745

ER -