Identification of oxidized proteins in rat plasma using avidin chromatography and tandem mass spectrometry

Hamid Mirzaei, Beatriz Baena, Coral Barbas, Fred E. Regnier

Research output: Contribution to journalArticle

40 Citations (Scopus)

Abstract

The objective in much of the proteomics literature today is to establish the difference between healthy and disease states at the protein level using blood plasma. A critical component in this endeavor is to establish what is normal. The focus of the work reported here was to do this with oxidized proteins that might relate to oxidative stress and oxidative stress-related diseases. Oxidative stress is known to increase markedly in cancer, diabetes, heart disease, and neurodegenerative diseases. Since proteins are one of the targets of ROS, generated by oxidative stress, oxidized proteins are excellent biomarker candidates for these diseases. But first it is necessary to identify oxidized proteins that occur in the healthy state. Healthy rat plasma was used in this study as a source for the identification of naturally oxidized proteins. Freshly drawn blood was treated with biotin hydrazide to selectively derivatize carbonyl groups in oxidized proteins. Oxidized proteins thus biotinylated were separated from the other plasma proteins using avidin affinity chromatography. Affinity selected proteins were further fractionated on a C8 RP column and fractions collected. The collected fractions were then tryptic digested and the peptides identified using a combination of LC/MS/MS and database searches. One hundred forty-six proteins were identified using 700 signature peptides from the tryptic digested chromatographic fractions. The most frequently encountered proteins in the samples were keratins. Brain and liver were among the organs contributing the most oxidized proteins to plasma followed by heart and kidney.

Original languageEnglish (US)
Pages (from-to)1516-1527
Number of pages12
JournalProteomics
Volume8
Issue number7
DOIs
StatePublished - Apr 2008

Fingerprint

Avidin
Tandem Mass Spectrometry
Chromatography
Mass spectrometry
Rats
Plasmas
Proteins
Oxidative stress
Oxidative Stress
Blood Proteins
Blood
Peptides
Neurodegenerative diseases
Affinity chromatography
Keratins
Heat-Shock Proteins
Affinity Chromatography
Neurodegenerative Diseases
Proteomics
Biomarkers

Keywords

  • Carbonylated proteins
  • Carbonylation
  • Oxidized keratin
  • Plasma oxidized proteins

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Identification of oxidized proteins in rat plasma using avidin chromatography and tandem mass spectrometry. / Mirzaei, Hamid; Baena, Beatriz; Barbas, Coral; Regnier, Fred E.

In: Proteomics, Vol. 8, No. 7, 04.2008, p. 1516-1527.

Research output: Contribution to journalArticle

Mirzaei, Hamid ; Baena, Beatriz ; Barbas, Coral ; Regnier, Fred E. / Identification of oxidized proteins in rat plasma using avidin chromatography and tandem mass spectrometry. In: Proteomics. 2008 ; Vol. 8, No. 7. pp. 1516-1527.
@article{1637d351e7fa400aaaf4b7eca1301b61,
title = "Identification of oxidized proteins in rat plasma using avidin chromatography and tandem mass spectrometry",
abstract = "The objective in much of the proteomics literature today is to establish the difference between healthy and disease states at the protein level using blood plasma. A critical component in this endeavor is to establish what is normal. The focus of the work reported here was to do this with oxidized proteins that might relate to oxidative stress and oxidative stress-related diseases. Oxidative stress is known to increase markedly in cancer, diabetes, heart disease, and neurodegenerative diseases. Since proteins are one of the targets of ROS, generated by oxidative stress, oxidized proteins are excellent biomarker candidates for these diseases. But first it is necessary to identify oxidized proteins that occur in the healthy state. Healthy rat plasma was used in this study as a source for the identification of naturally oxidized proteins. Freshly drawn blood was treated with biotin hydrazide to selectively derivatize carbonyl groups in oxidized proteins. Oxidized proteins thus biotinylated were separated from the other plasma proteins using avidin affinity chromatography. Affinity selected proteins were further fractionated on a C8 RP column and fractions collected. The collected fractions were then tryptic digested and the peptides identified using a combination of LC/MS/MS and database searches. One hundred forty-six proteins were identified using 700 signature peptides from the tryptic digested chromatographic fractions. The most frequently encountered proteins in the samples were keratins. Brain and liver were among the organs contributing the most oxidized proteins to plasma followed by heart and kidney.",
keywords = "Carbonylated proteins, Carbonylation, Oxidized keratin, Plasma oxidized proteins",
author = "Hamid Mirzaei and Beatriz Baena and Coral Barbas and Regnier, {Fred E.}",
year = "2008",
month = "4",
doi = "10.1002/pmic.200700363",
language = "English (US)",
volume = "8",
pages = "1516--1527",
journal = "Proteomics",
issn = "1615-9853",
publisher = "Wiley-VCH Verlag",
number = "7",

}

TY - JOUR

T1 - Identification of oxidized proteins in rat plasma using avidin chromatography and tandem mass spectrometry

AU - Mirzaei, Hamid

AU - Baena, Beatriz

AU - Barbas, Coral

AU - Regnier, Fred E.

PY - 2008/4

Y1 - 2008/4

N2 - The objective in much of the proteomics literature today is to establish the difference between healthy and disease states at the protein level using blood plasma. A critical component in this endeavor is to establish what is normal. The focus of the work reported here was to do this with oxidized proteins that might relate to oxidative stress and oxidative stress-related diseases. Oxidative stress is known to increase markedly in cancer, diabetes, heart disease, and neurodegenerative diseases. Since proteins are one of the targets of ROS, generated by oxidative stress, oxidized proteins are excellent biomarker candidates for these diseases. But first it is necessary to identify oxidized proteins that occur in the healthy state. Healthy rat plasma was used in this study as a source for the identification of naturally oxidized proteins. Freshly drawn blood was treated with biotin hydrazide to selectively derivatize carbonyl groups in oxidized proteins. Oxidized proteins thus biotinylated were separated from the other plasma proteins using avidin affinity chromatography. Affinity selected proteins were further fractionated on a C8 RP column and fractions collected. The collected fractions were then tryptic digested and the peptides identified using a combination of LC/MS/MS and database searches. One hundred forty-six proteins were identified using 700 signature peptides from the tryptic digested chromatographic fractions. The most frequently encountered proteins in the samples were keratins. Brain and liver were among the organs contributing the most oxidized proteins to plasma followed by heart and kidney.

AB - The objective in much of the proteomics literature today is to establish the difference between healthy and disease states at the protein level using blood plasma. A critical component in this endeavor is to establish what is normal. The focus of the work reported here was to do this with oxidized proteins that might relate to oxidative stress and oxidative stress-related diseases. Oxidative stress is known to increase markedly in cancer, diabetes, heart disease, and neurodegenerative diseases. Since proteins are one of the targets of ROS, generated by oxidative stress, oxidized proteins are excellent biomarker candidates for these diseases. But first it is necessary to identify oxidized proteins that occur in the healthy state. Healthy rat plasma was used in this study as a source for the identification of naturally oxidized proteins. Freshly drawn blood was treated with biotin hydrazide to selectively derivatize carbonyl groups in oxidized proteins. Oxidized proteins thus biotinylated were separated from the other plasma proteins using avidin affinity chromatography. Affinity selected proteins were further fractionated on a C8 RP column and fractions collected. The collected fractions were then tryptic digested and the peptides identified using a combination of LC/MS/MS and database searches. One hundred forty-six proteins were identified using 700 signature peptides from the tryptic digested chromatographic fractions. The most frequently encountered proteins in the samples were keratins. Brain and liver were among the organs contributing the most oxidized proteins to plasma followed by heart and kidney.

KW - Carbonylated proteins

KW - Carbonylation

KW - Oxidized keratin

KW - Plasma oxidized proteins

UR - http://www.scopus.com/inward/record.url?scp=42049099009&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=42049099009&partnerID=8YFLogxK

U2 - 10.1002/pmic.200700363

DO - 10.1002/pmic.200700363

M3 - Article

C2 - 18383005

AN - SCOPUS:42049099009

VL - 8

SP - 1516

EP - 1527

JO - Proteomics

JF - Proteomics

SN - 1615-9853

IS - 7

ER -