Quantitative analysis of modified proteins and their positional isomers by tandem mass spectrometry: Human histone H4

James J. Pesavento, Craig Andrew Mizzen, Neil L. Kelleher

Research output: Contribution to journalArticle

Abstract

Here we show that fragment ion abundances from dissociation of ions created from mixtures of multiply modified histone H4 (11 kDa) or of N-terminal synthetic peptides (2 kDa) correspond to their respective intact ion abundances measured by Fourier transform mass spectrometry. Isomeric mixtures of modified forms of the same protein are resolved and quantitated with a precision of ≤5% using the relative ratios of their fragment ions, with intact protein ions created by electrospray greatly easing many of the systematic biases that more strongly affect small peptides (e.g., differences in ionization efficiency and ion m/z values). The ion fragmentation methods validated here are directly extensible to intact human proteins to derive quantitative information on the highly related and often isomeric protein forms created by combinatorial arrays of posttranslational modifications.

Original languageEnglish (US)
Pages (from-to)4271-4280
Number of pages10
JournalAnalytical chemistry
Volume78
Issue number13
DOIs
StatePublished - Jul 1 2006

Fingerprint

Isomers
Histones
Mass spectrometry
Ions
Chemical analysis
Proteins
Peptides
Ionization
Fourier transforms

ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

Quantitative analysis of modified proteins and their positional isomers by tandem mass spectrometry : Human histone H4. / Pesavento, James J.; Mizzen, Craig Andrew; Kelleher, Neil L.

In: Analytical chemistry, Vol. 78, No. 13, 01.07.2006, p. 4271-4280.

Research output: Contribution to journalArticle

Pesavento, James J. ; Mizzen, Craig Andrew ; Kelleher, Neil L. / Quantitative analysis of modified proteins and their positional isomers by tandem mass spectrometry : Human histone H4. In: Analytical chemistry. 2006 ; Vol. 78, No. 13. pp. 4271-4280.
@article{bdb03829008a4066b5b7b7100a37e703,
title = "Quantitative analysis of modified proteins and their positional isomers by tandem mass spectrometry: Human histone H4",
abstract = "Here we show that fragment ion abundances from dissociation of ions created from mixtures of multiply modified histone H4 (11 kDa) or of N-terminal synthetic peptides (2 kDa) correspond to their respective intact ion abundances measured by Fourier transform mass spectrometry. Isomeric mixtures of modified forms of the same protein are resolved and quantitated with a precision of ≤5{\%} using the relative ratios of their fragment ions, with intact protein ions created by electrospray greatly easing many of the systematic biases that more strongly affect small peptides (e.g., differences in ionization efficiency and ion m/z values). The ion fragmentation methods validated here are directly extensible to intact human proteins to derive quantitative information on the highly related and often isomeric protein forms created by combinatorial arrays of posttranslational modifications.",
author = "Pesavento, {James J.} and Mizzen, {Craig Andrew} and Kelleher, {Neil L.}",
year = "2006",
month = "7",
day = "1",
doi = "10.1021/ac0600050",
language = "English (US)",
volume = "78",
pages = "4271--4280",
journal = "Analytical Chemistry",
issn = "0003-2700",
publisher = "American Chemical Society",
number = "13",

}

TY - JOUR

T1 - Quantitative analysis of modified proteins and their positional isomers by tandem mass spectrometry

T2 - Human histone H4

AU - Pesavento, James J.

AU - Mizzen, Craig Andrew

AU - Kelleher, Neil L.

PY - 2006/7/1

Y1 - 2006/7/1

N2 - Here we show that fragment ion abundances from dissociation of ions created from mixtures of multiply modified histone H4 (11 kDa) or of N-terminal synthetic peptides (2 kDa) correspond to their respective intact ion abundances measured by Fourier transform mass spectrometry. Isomeric mixtures of modified forms of the same protein are resolved and quantitated with a precision of ≤5% using the relative ratios of their fragment ions, with intact protein ions created by electrospray greatly easing many of the systematic biases that more strongly affect small peptides (e.g., differences in ionization efficiency and ion m/z values). The ion fragmentation methods validated here are directly extensible to intact human proteins to derive quantitative information on the highly related and often isomeric protein forms created by combinatorial arrays of posttranslational modifications.

AB - Here we show that fragment ion abundances from dissociation of ions created from mixtures of multiply modified histone H4 (11 kDa) or of N-terminal synthetic peptides (2 kDa) correspond to their respective intact ion abundances measured by Fourier transform mass spectrometry. Isomeric mixtures of modified forms of the same protein are resolved and quantitated with a precision of ≤5% using the relative ratios of their fragment ions, with intact protein ions created by electrospray greatly easing many of the systematic biases that more strongly affect small peptides (e.g., differences in ionization efficiency and ion m/z values). The ion fragmentation methods validated here are directly extensible to intact human proteins to derive quantitative information on the highly related and often isomeric protein forms created by combinatorial arrays of posttranslational modifications.

UR - http://www.scopus.com/inward/record.url?scp=33745700686&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33745700686&partnerID=8YFLogxK

U2 - 10.1021/ac0600050

DO - 10.1021/ac0600050

M3 - Article

C2 - 16808433

AN - SCOPUS:33745700686

VL - 78

SP - 4271

EP - 4280

JO - Analytical Chemistry

JF - Analytical Chemistry

SN - 0003-2700

IS - 13

ER -