Proteomic investigation of natural killer cell microsomes using gas-phase fractionation by mass spectrometry

Josip Blonder; Maria Cecilia Rodriguez-Galan; David A. Lucas; Howard A. Young; Haleem J. Issaq; Timothy D. Veenstra; Thomas P. Conrads

doi:10.1016/j.bbapap.2003.10.009

Proteomic investigation of natural killer cell microsomes using gas-phase fractionation by mass spectrometry

Josip Blonder, Maria Cecilia Rodriguez-Galan, David A. Lucas, Howard A. Young, Haleem J. Issaq, Timothy D. Veenstra, Thomas P. Conrads^*

^*Corresponding author for this work

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

25 Scopus citations

Abstract

We have explored the utility of gas-phase fractionation by mass spectrometry (MS) in the mass-to-charge (m/z) dimension (GPF_m/z) for increasing the effective number of protein identifications in cases where sample quantity limits the use of multi-dimensional chromatographic fractionation. A peptide digestate from proteins isolated from the membrane fraction of natural killer (NK) cells was analyzed by microcapillary reversed-phase liquid chromatography coupled online to an ion-trap (IT) mass spectrometer. Performing GPF_m/z using eight narrow precursor ion scan m/z ranges enabled the identification of 340 NK cell proteins from 12 μg of digestate, representing more than a fivefold increase in the number of proteins identified as compared to the same experiment employing a standard precursor ion survey scan m/z range (i.e., m/z 400-2000). The results show that GPF_m/z represents an effective technique for increasing protein identifications in global proteomic investigations especially when sample quantity is limited.

Original language	English
Pages (from-to)	87-95
Number of pages	9
Journal	Biochimica et Biophysica Acta - Proteins and Proteomics
Volume	1698
Issue number	1
DOIs	https://doi.org/10.1016/j.bbapap.2003.10.009
State	Published - 8 Apr 2004
Externally published	Yes

Keywords

2D PAGE
Gas-phase fractionation
MS
MS/MS
Mass spectrometry
Membrane proteomic
Microcapillary liquid chromatography
Microsome
Natural killer cell
Tandem mass spectrometry
Two-dimensional polyacrylamide gel electrophoresis
m/z
μLC

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10.1016/j.bbapap.2003.10.009

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title = "Proteomic investigation of natural killer cell microsomes using gas-phase fractionation by mass spectrometry",

abstract = "We have explored the utility of gas-phase fractionation by mass spectrometry (MS) in the mass-to-charge (m/z) dimension (GPFm/z) for increasing the effective number of protein identifications in cases where sample quantity limits the use of multi-dimensional chromatographic fractionation. A peptide digestate from proteins isolated from the membrane fraction of natural killer (NK) cells was analyzed by microcapillary reversed-phase liquid chromatography coupled online to an ion-trap (IT) mass spectrometer. Performing GPFm/z using eight narrow precursor ion scan m/z ranges enabled the identification of 340 NK cell proteins from 12 μg of digestate, representing more than a fivefold increase in the number of proteins identified as compared to the same experiment employing a standard precursor ion survey scan m/z range (i.e., m/z 400-2000). The results show that GPFm/z represents an effective technique for increasing protein identifications in global proteomic investigations especially when sample quantity is limited.",

keywords = "2D PAGE, Gas-phase fractionation, MS, MS/MS, Mass spectrometry, Membrane proteomic, Microcapillary liquid chromatography, Microsome, Natural killer cell, Tandem mass spectrometry, Two-dimensional polyacrylamide gel electrophoresis, m/z, μLC",

author = "Josip Blonder and Rodriguez-Galan, {Maria Cecilia} and Lucas, {David A.} and Young, {Howard A.} and Issaq, {Haleem J.} and Veenstra, {Timothy D.} and Conrads, {Thomas P.}",

note = "Funding Information: By acceptance of this article, the publisher or recipient acknowledges the right of the United States Government to retain a nonexclusive, royalty-free license and to any copyright covering the article. The content of this publication does not necessarily reflect the views or policies of the Department of Health and Human Services, nor does mention of trade names, commercial products, or organization imply endorsement by the United States Government. This project has been funded in whole or in part with Federal funds from the National Cancer Institute, National Institutes of Health, under contract NO1-CO-12400.",

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doi = "10.1016/j.bbapap.2003.10.009",

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AU - Blonder, Josip

AU - Rodriguez-Galan, Maria Cecilia

AU - Lucas, David A.

AU - Young, Howard A.

AU - Issaq, Haleem J.

AU - Veenstra, Timothy D.

AU - Conrads, Thomas P.

N1 - Funding Information: By acceptance of this article, the publisher or recipient acknowledges the right of the United States Government to retain a nonexclusive, royalty-free license and to any copyright covering the article. The content of this publication does not necessarily reflect the views or policies of the Department of Health and Human Services, nor does mention of trade names, commercial products, or organization imply endorsement by the United States Government. This project has been funded in whole or in part with Federal funds from the National Cancer Institute, National Institutes of Health, under contract NO1-CO-12400.

PY - 2004/4/8

Y1 - 2004/4/8

N2 - We have explored the utility of gas-phase fractionation by mass spectrometry (MS) in the mass-to-charge (m/z) dimension (GPFm/z) for increasing the effective number of protein identifications in cases where sample quantity limits the use of multi-dimensional chromatographic fractionation. A peptide digestate from proteins isolated from the membrane fraction of natural killer (NK) cells was analyzed by microcapillary reversed-phase liquid chromatography coupled online to an ion-trap (IT) mass spectrometer. Performing GPFm/z using eight narrow precursor ion scan m/z ranges enabled the identification of 340 NK cell proteins from 12 μg of digestate, representing more than a fivefold increase in the number of proteins identified as compared to the same experiment employing a standard precursor ion survey scan m/z range (i.e., m/z 400-2000). The results show that GPFm/z represents an effective technique for increasing protein identifications in global proteomic investigations especially when sample quantity is limited.

AB - We have explored the utility of gas-phase fractionation by mass spectrometry (MS) in the mass-to-charge (m/z) dimension (GPFm/z) for increasing the effective number of protein identifications in cases where sample quantity limits the use of multi-dimensional chromatographic fractionation. A peptide digestate from proteins isolated from the membrane fraction of natural killer (NK) cells was analyzed by microcapillary reversed-phase liquid chromatography coupled online to an ion-trap (IT) mass spectrometer. Performing GPFm/z using eight narrow precursor ion scan m/z ranges enabled the identification of 340 NK cell proteins from 12 μg of digestate, representing more than a fivefold increase in the number of proteins identified as compared to the same experiment employing a standard precursor ion survey scan m/z range (i.e., m/z 400-2000). The results show that GPFm/z represents an effective technique for increasing protein identifications in global proteomic investigations especially when sample quantity is limited.

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KW - Microsome

KW - Natural killer cell

KW - Tandem mass spectrometry

KW - Two-dimensional polyacrylamide gel electrophoresis

KW - m/z

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JO - Biochimica et Biophysica Acta - Proteins and Proteomics

JF - Biochimica et Biophysica Acta - Proteins and Proteomics

IS - 1

ER -

Proteomic investigation of natural killer cell microsomes using gas-phase fractionation by mass spectrometry

Abstract

Keywords

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