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Absolute SILAC-compatible expression strain allows SUMO-2 copy number determination in clinical samples

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Absolute SILAC-compatible expression strain allows SUMO-2 copy number determination in clinical samples. / Matic, Ivan; Jaffray, Ellis G.; Oxenham, Senga K.; Groves, Michael J.; Barratt, Christopher L. R.; Tauro, Sudhir; Stanley-Wall, Nicola R.; Hay, Ronald T.

In: Journal of Proteome Research, Vol. 10, No. 10, 10.2011, p. 4869-4875.

Research output: Contribution to journalArticle

Harvard

Matic, I, Jaffray, EG, Oxenham, SK, Groves, MJ, Barratt, CLR, Tauro, S, Stanley-Wall, NR & Hay, RT 2011, 'Absolute SILAC-compatible expression strain allows SUMO-2 copy number determination in clinical samples' Journal of Proteome Research, vol 10, no. 10, pp. 4869-4875., 10.1021/pr2004715

APA

Matic, I., Jaffray, E. G., Oxenham, S. K., Groves, M. J., Barratt, C. L. R., Tauro, S., ... Hay, R. T. (2011). Absolute SILAC-compatible expression strain allows SUMO-2 copy number determination in clinical samples. Journal of Proteome Research, 10(10), 4869-4875. 10.1021/pr2004715

Vancouver

Matic I, Jaffray EG, Oxenham SK, Groves MJ, Barratt CLR, Tauro S et al. Absolute SILAC-compatible expression strain allows SUMO-2 copy number determination in clinical samples. Journal of Proteome Research. 2011 Oct;10(10):4869-4875. Available from: 10.1021/pr2004715

Author

Matic, Ivan; Jaffray, Ellis G.; Oxenham, Senga K.; Groves, Michael J.; Barratt, Christopher L. R.; Tauro, Sudhir; Stanley-Wall, Nicola R.; Hay, Ronald T. / Absolute SILAC-compatible expression strain allows SUMO-2 copy number determination in clinical samples.

In: Journal of Proteome Research, Vol. 10, No. 10, 10.2011, p. 4869-4875.

Research output: Contribution to journalArticle

Bibtex - Download

@article{3f0731c7326e46aab80c7978c8f58615,
title = "Absolute SILAC-compatible expression strain allows SUMO-2 copy number determination in clinical samples",
keywords = "Absolute SILAC, PSAQ, SUMO, Quantitation, Mass spectrometry, Proteomics, Biomarker, Sperm cells, Chronic lymphocytic leukemia, Spectrometry based proteomics, Cell culture SILAC, Quantitative proteomics, Amino acids, Protein quantification, Identification, Fluorescent, Standards, Peptides, Genes",
author = "Ivan Matic and Jaffray, {Ellis G.} and Oxenham, {Senga K.} and Groves, {Michael J.} and Barratt, {Christopher L. R.} and Sudhir Tauro and Stanley-Wall, {Nicola R.} and Hay, {Ronald T.}",
year = "2011",
doi = "10.1021/pr2004715",
volume = "10",
number = "10",
pages = "4869--4875",
journal = "Journal of Proteome Research",
issn = "1535-3893",

}

RIS (suitable for import to EndNote) - Download

TY - JOUR

T1 - Absolute SILAC-compatible expression strain allows SUMO-2 copy number determination in clinical samples

A1 - Matic,Ivan

A1 - Jaffray,Ellis G.

A1 - Oxenham,Senga K.

A1 - Groves,Michael J.

A1 - Barratt,Christopher L. R.

A1 - Tauro,Sudhir

A1 - Stanley-Wall,Nicola R.

A1 - Hay,Ronald T.

AU - Matic,Ivan

AU - Jaffray,Ellis G.

AU - Oxenham,Senga K.

AU - Groves,Michael J.

AU - Barratt,Christopher L. R.

AU - Tauro,Sudhir

AU - Stanley-Wall,Nicola R.

AU - Hay,Ronald T.

PY - 2011/10

Y1 - 2011/10

N2 - <p>Quantitative mass spectrometry-based proteomics is a vital tool in modern life science research. In contrast to the popularity of approaches for relative protein quantitation, the widespread use of absolute quantitation has been hampered by inefficient and expensive production of labeled protein standards. To optimize production of isotopically labeled standards, we genetically modified a commonly employed protein expression Escherichia coil strain, BL21 (DE3), to construct an auxotroph for arginine and lysine. This bacterial strain allows low-cost, high-level expression of fully labeled proteins with no conversion of labeled arginine to proline. In combination with a fluorescence-based quantitation of standards and nontargeted LC-MS/MS analysis of unfractionated total cell lysates, this strain was used to determine the copy number of a post-translational modifier, small ubiquitin-like modifier (SUMO-2), in HeLa, human sperm, and chronic lymphocytic leukemia cells. By streamlining and improving the generation of labeled standards, this production system increases the breadth of absolute quantitation by mass spectrometry and will facilitate a far wider uptake of this important technique than previously possible.</p>

AB - <p>Quantitative mass spectrometry-based proteomics is a vital tool in modern life science research. In contrast to the popularity of approaches for relative protein quantitation, the widespread use of absolute quantitation has been hampered by inefficient and expensive production of labeled protein standards. To optimize production of isotopically labeled standards, we genetically modified a commonly employed protein expression Escherichia coil strain, BL21 (DE3), to construct an auxotroph for arginine and lysine. This bacterial strain allows low-cost, high-level expression of fully labeled proteins with no conversion of labeled arginine to proline. In combination with a fluorescence-based quantitation of standards and nontargeted LC-MS/MS analysis of unfractionated total cell lysates, this strain was used to determine the copy number of a post-translational modifier, small ubiquitin-like modifier (SUMO-2), in HeLa, human sperm, and chronic lymphocytic leukemia cells. By streamlining and improving the generation of labeled standards, this production system increases the breadth of absolute quantitation by mass spectrometry and will facilitate a far wider uptake of this important technique than previously possible.</p>

KW - Absolute SILAC

KW - PSAQ

KW - SUMO

KW - Quantitation

KW - Mass spectrometry

KW - Proteomics

KW - Biomarker

KW - Sperm cells

KW - Chronic lymphocytic leukemia

KW - Spectrometry based proteomics

KW - Cell culture SILAC

KW - Quantitative proteomics

KW - Amino acids

KW - Protein quantification

KW - Identification

KW - Fluorescent

KW - Standards

KW - Peptides

KW - Genes

U2 - 10.1021/pr2004715

DO - 10.1021/pr2004715

M1 - Article

JO - Journal of Proteome Research

JF - Journal of Proteome Research

SN - 1535-3893

IS - 10

VL - 10

SP - 4869

EP - 4875

ER -

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