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Application of displacement chromatography for the analysis of a lipid raft proteome

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Application of displacement chromatography for the analysis of a lipid raft proteome. / Trusch, Maria; Böhlick, Alexandra; Hildebrand, Diana; Lichtner, Björn; Bertsch, Andreas; Kohlbacher, Oliver; Bachmann, Sebastian; Schlüter, Hartmut.

In: J CHROMATOGR B, Vol. 878, No. 3-4, 01.02.2010, p. 309-14.

Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review

Harvard

Trusch, M, Böhlick, A, Hildebrand, D, Lichtner, B, Bertsch, A, Kohlbacher, O, Bachmann, S & Schlüter, H 2010, 'Application of displacement chromatography for the analysis of a lipid raft proteome', J CHROMATOGR B, vol. 878, no. 3-4, pp. 309-14. https://doi.org/10.1016/j.jchromb.2009.11.035

APA

Trusch, M., Böhlick, A., Hildebrand, D., Lichtner, B., Bertsch, A., Kohlbacher, O., Bachmann, S., & Schlüter, H. (2010). Application of displacement chromatography for the analysis of a lipid raft proteome. J CHROMATOGR B, 878(3-4), 309-14. https://doi.org/10.1016/j.jchromb.2009.11.035

Vancouver

Trusch M, Böhlick A, Hildebrand D, Lichtner B, Bertsch A, Kohlbacher O et al. Application of displacement chromatography for the analysis of a lipid raft proteome. J CHROMATOGR B. 2010 Feb 1;878(3-4):309-14. https://doi.org/10.1016/j.jchromb.2009.11.035

Bibtex

@article{664251ac5baf40718bf5c5977ac084a2,
title = "Application of displacement chromatography for the analysis of a lipid raft proteome",
abstract = "Defining membrane proteomes is fundamental to understand the role of membrane proteins in biological processes and to find new targets for drug development. Usually multidimensional chromatography using step or gradient elution is applied for the separation of tryptic peptides of membrane proteins prior to their mass spectrometric analysis. Displacement chromatography (DC) offers several advantages that are helpful for proteome analysis. However, DC has so far been applied for proteomic investigations only in few cases. In this study we therefore applied DC in a multidimensional LC-MS approach for the separation and identification of membrane proteins located in cholesterol-enriched membrane microdomains (lipid rafts) obtained from rat kidney by density gradient centrifugation. The tryptic peptides were separated on a cation-exchange column in the displacement mode with spermine used as displacer. Fractions obtained from DC were analyzed using an HPLC-chip system coupled to an electrospray-ionization ion-trap mass spectrometer. This procedure yielded more than 400 highly significant peptide spectrum matches and led to the identification of more than 140 reliable protein hits within an established rat kidney lipid raft proteome. The majority of identified proteins were membrane proteins. In sum, our results demonstrate that DC is a suitable alternative to gradient elution separations for the identification of proteins via a multidimensional LC-MS approach.",
keywords = "Amino Acid Sequence, Animals, Cation Exchange Resins, Chromatography, Liquid, Immunoblotting, Kidney, Male, Mass Spectrometry, Membrane Microdomains, Molecular Sequence Data, Peptides, Proteome, Rats, Rats, Wistar",
author = "Maria Trusch and Alexandra B{\"o}hlick and Diana Hildebrand and Bj{\"o}rn Lichtner and Andreas Bertsch and Oliver Kohlbacher and Sebastian Bachmann and Hartmut Schl{\"u}ter",
note = "2009 Elsevier B.V. All rights reserved.",
year = "2010",
month = feb,
day = "1",
doi = "10.1016/j.jchromb.2009.11.035",
language = "English",
volume = "878",
pages = "309--14",
journal = "J CHROMATOGR B",
issn = "1570-0232",
publisher = "Elsevier",
number = "3-4",

}

RIS

TY - JOUR

T1 - Application of displacement chromatography for the analysis of a lipid raft proteome

AU - Trusch, Maria

AU - Böhlick, Alexandra

AU - Hildebrand, Diana

AU - Lichtner, Björn

AU - Bertsch, Andreas

AU - Kohlbacher, Oliver

AU - Bachmann, Sebastian

AU - Schlüter, Hartmut

N1 - 2009 Elsevier B.V. All rights reserved.

PY - 2010/2/1

Y1 - 2010/2/1

N2 - Defining membrane proteomes is fundamental to understand the role of membrane proteins in biological processes and to find new targets for drug development. Usually multidimensional chromatography using step or gradient elution is applied for the separation of tryptic peptides of membrane proteins prior to their mass spectrometric analysis. Displacement chromatography (DC) offers several advantages that are helpful for proteome analysis. However, DC has so far been applied for proteomic investigations only in few cases. In this study we therefore applied DC in a multidimensional LC-MS approach for the separation and identification of membrane proteins located in cholesterol-enriched membrane microdomains (lipid rafts) obtained from rat kidney by density gradient centrifugation. The tryptic peptides were separated on a cation-exchange column in the displacement mode with spermine used as displacer. Fractions obtained from DC were analyzed using an HPLC-chip system coupled to an electrospray-ionization ion-trap mass spectrometer. This procedure yielded more than 400 highly significant peptide spectrum matches and led to the identification of more than 140 reliable protein hits within an established rat kidney lipid raft proteome. The majority of identified proteins were membrane proteins. In sum, our results demonstrate that DC is a suitable alternative to gradient elution separations for the identification of proteins via a multidimensional LC-MS approach.

AB - Defining membrane proteomes is fundamental to understand the role of membrane proteins in biological processes and to find new targets for drug development. Usually multidimensional chromatography using step or gradient elution is applied for the separation of tryptic peptides of membrane proteins prior to their mass spectrometric analysis. Displacement chromatography (DC) offers several advantages that are helpful for proteome analysis. However, DC has so far been applied for proteomic investigations only in few cases. In this study we therefore applied DC in a multidimensional LC-MS approach for the separation and identification of membrane proteins located in cholesterol-enriched membrane microdomains (lipid rafts) obtained from rat kidney by density gradient centrifugation. The tryptic peptides were separated on a cation-exchange column in the displacement mode with spermine used as displacer. Fractions obtained from DC were analyzed using an HPLC-chip system coupled to an electrospray-ionization ion-trap mass spectrometer. This procedure yielded more than 400 highly significant peptide spectrum matches and led to the identification of more than 140 reliable protein hits within an established rat kidney lipid raft proteome. The majority of identified proteins were membrane proteins. In sum, our results demonstrate that DC is a suitable alternative to gradient elution separations for the identification of proteins via a multidimensional LC-MS approach.

KW - Amino Acid Sequence

KW - Animals

KW - Cation Exchange Resins

KW - Chromatography, Liquid

KW - Immunoblotting

KW - Kidney

KW - Male

KW - Mass Spectrometry

KW - Membrane Microdomains

KW - Molecular Sequence Data

KW - Peptides

KW - Proteome

KW - Rats

KW - Rats, Wistar

U2 - 10.1016/j.jchromb.2009.11.035

DO - 10.1016/j.jchromb.2009.11.035

M3 - SCORING: Journal article

C2 - 20015709

VL - 878

SP - 309

EP - 314

JO - J CHROMATOGR B

JF - J CHROMATOGR B

SN - 1570-0232

IS - 3-4

ER -