Alternative fusion protein strategies to express recalcitrant QconCAT proteins for quantitative proteomics of human drug metabolizing enzymes and transporters

Matthew R. Russell; Brahim Achour; Edward A. McKenzie; Ruth Lopez; Matthew D. Harwood; Amin Rostami-Hodjegan; Jill Barber

doi:10.1021/pr400279u

Alternative fusion protein strategies to express recalcitrant QconCAT proteins for quantitative proteomics of human drug metabolizing enzymes and transporters

Matthew R. Russell, Brahim Achour, Edward A. McKenzie, Ruth Lopez, Matthew D. Harwood, Amin Rostami-Hodjegan, Jill Barber

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

Abstract

QconCAT is a tool for quantitative proteomics, consisting of an artificial protein, expressed from an artificial gene, made up of a concatenated string of proteotypic peptides selected from the proteins under study. Isotopically labeled QconCAT (usually containing 13C6-arginine and 13C6-lysine) provides a standard for each proteotypic peptide included in its sequence. In practice, some QconCAT proteins fail to express at sufficient levels for the purpose of quantitative analysis. Two complementary methods are presented to express recalcitrant QconCAT proteins intended to quantify human hepatic enzymes and transporters. © 2013 American Chemical Society.

Original language	English
Pages (from-to)	5934-5942
Number of pages	8
Journal	Journal of Proteome Research
Volume	12
Issue number	12
DOIs	https://doi.org/10.1021/pr400279u
Publication status	Published - 6 Dec 2013

Keywords

fusion construct
insoluble fusion expression protein
MetCAT
NUSa-soluble fusion expression protein
SDS-PAGE
TransCAT
triple-quadrupole mass spectrometry

Research Beacons, Institutes and Platforms

Manchester Institute of Biotechnology

Access to Document

10.1021/pr400279u

Biological Mass Spectrometry (BioMS) Facility
Knight, D. (Core Facility Lead), Warwood, S. (Senior Technical Specialist), Selley, J. (Technical Specialist), Taylor, G. (Technical Specialist), Fullwood, P. (Technical Specialist), Keevill, E.-J. (Senior Technician) & Allsey, J. (Technician)
FBMH Platform Sciences, Enabling Technologies & Infrastructure
Facility/equipment: Facility

Cite this

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title = "Alternative fusion protein strategies to express recalcitrant QconCAT proteins for quantitative proteomics of human drug metabolizing enzymes and transporters",

abstract = "QconCAT is a tool for quantitative proteomics, consisting of an artificial protein, expressed from an artificial gene, made up of a concatenated string of proteotypic peptides selected from the proteins under study. Isotopically labeled QconCAT (usually containing 13C6-arginine and 13C6-lysine) provides a standard for each proteotypic peptide included in its sequence. In practice, some QconCAT proteins fail to express at sufficient levels for the purpose of quantitative analysis. Two complementary methods are presented to express recalcitrant QconCAT proteins intended to quantify human hepatic enzymes and transporters. {\textcopyright} 2013 American Chemical Society.",

keywords = "fusion construct, insoluble fusion expression protein, MetCAT, NUSa-soluble fusion expression protein, SDS-PAGE, TransCAT, triple-quadrupole mass spectrometry",

author = "Russell, \{Matthew R.\} and Brahim Achour and McKenzie, \{Edward A.\} and Ruth Lopez and Harwood, \{Matthew D.\} and Amin Rostami-Hodjegan and Jill Barber",

year = "2013",

month = dec,

day = "6",

doi = "10.1021/pr400279u",

language = "English",

volume = "12",

pages = "5934--5942",

journal = "Journal of Proteome Research",

issn = "1535-3893",

publisher = "American Chemical Society",

number = "12",

}

Alternative fusion protein strategies to express recalcitrant QconCAT proteins for quantitative proteomics of human drug metabolizing enzymes and transporters. / Russell, Matthew R.; Achour, Brahim; McKenzie, Edward A. et al.
In: Journal of Proteome Research, Vol. 12, No. 12, 06.12.2013, p. 5934-5942.

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

TY - JOUR

T1 - Alternative fusion protein strategies to express recalcitrant QconCAT proteins for quantitative proteomics of human drug metabolizing enzymes and transporters

AU - Russell, Matthew R.

AU - Achour, Brahim

AU - McKenzie, Edward A.

AU - Lopez, Ruth

AU - Harwood, Matthew D.

AU - Rostami-Hodjegan, Amin

AU - Barber, Jill

PY - 2013/12/6

Y1 - 2013/12/6

N2 - QconCAT is a tool for quantitative proteomics, consisting of an artificial protein, expressed from an artificial gene, made up of a concatenated string of proteotypic peptides selected from the proteins under study. Isotopically labeled QconCAT (usually containing 13C6-arginine and 13C6-lysine) provides a standard for each proteotypic peptide included in its sequence. In practice, some QconCAT proteins fail to express at sufficient levels for the purpose of quantitative analysis. Two complementary methods are presented to express recalcitrant QconCAT proteins intended to quantify human hepatic enzymes and transporters. © 2013 American Chemical Society.

AB - QconCAT is a tool for quantitative proteomics, consisting of an artificial protein, expressed from an artificial gene, made up of a concatenated string of proteotypic peptides selected from the proteins under study. Isotopically labeled QconCAT (usually containing 13C6-arginine and 13C6-lysine) provides a standard for each proteotypic peptide included in its sequence. In practice, some QconCAT proteins fail to express at sufficient levels for the purpose of quantitative analysis. Two complementary methods are presented to express recalcitrant QconCAT proteins intended to quantify human hepatic enzymes and transporters. © 2013 American Chemical Society.

KW - fusion construct

KW - insoluble fusion expression protein

KW - MetCAT

KW - NUSa-soluble fusion expression protein

KW - SDS-PAGE

KW - TransCAT

KW - triple-quadrupole mass spectrometry

U2 - 10.1021/pr400279u

DO - 10.1021/pr400279u

M3 - Article

SN - 1535-3893

VL - 12

SP - 5934

EP - 5942

JO - Journal of Proteome Research

JF - Journal of Proteome Research

IS - 12

ER -

Alternative fusion protein strategies to express recalcitrant QconCAT proteins for quantitative proteomics of human drug metabolizing enzymes and transporters

Abstract

Keywords

Research Beacons, Institutes and Platforms

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Equipment

Biological Mass Spectrometry (BioMS) Facility

Cite this