A targeted proteomics toolkit for high-throughput absolute quantification of Escherichia coli proteins

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

Standard

A targeted proteomics toolkit for high-throughput absolute quantification of Escherichia coli proteins. / Batth, Tanveer S; Singh, Pragya; Ramakrishnan, Vikram R; Sousa, Mirta M L; Chan, Leanne Jade G; Tran, Huu M; Luning, Eric G; Pan, Eva H Y; Vuu, Khanh M; Keasling, Jay D; Adams, Paul D; Petzold, Christopher J.

In: Metabolic Engineering, Vol. 26, 2014, p. 48-56.

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

Harvard

Batth, TS, Singh, P, Ramakrishnan, VR, Sousa, MML, Chan, LJG, Tran, HM, Luning, EG, Pan, EHY, Vuu, KM, Keasling, JD, Adams, PD & Petzold, CJ 2014, 'A targeted proteomics toolkit for high-throughput absolute quantification of Escherichia coli proteins', Metabolic Engineering, vol. 26, pp. 48-56. https://doi.org/10.1016/j.ymben.2014.08.004

APA

Batth, T. S., Singh, P., Ramakrishnan, V. R., Sousa, M. M. L., Chan, L. J. G., Tran, H. M., Luning, E. G., Pan, E. H. Y., Vuu, K. M., Keasling, J. D., Adams, P. D., & Petzold, C. J. (2014). A targeted proteomics toolkit for high-throughput absolute quantification of Escherichia coli proteins. Metabolic Engineering, 26, 48-56. https://doi.org/10.1016/j.ymben.2014.08.004

Vancouver

Batth TS, Singh P, Ramakrishnan VR, Sousa MML, Chan LJG, Tran HM et al. A targeted proteomics toolkit for high-throughput absolute quantification of Escherichia coli proteins. Metabolic Engineering. 2014;26:48-56. https://doi.org/10.1016/j.ymben.2014.08.004

Author

Batth, Tanveer S ; Singh, Pragya ; Ramakrishnan, Vikram R ; Sousa, Mirta M L ; Chan, Leanne Jade G ; Tran, Huu M ; Luning, Eric G ; Pan, Eva H Y ; Vuu, Khanh M ; Keasling, Jay D ; Adams, Paul D ; Petzold, Christopher J. / A targeted proteomics toolkit for high-throughput absolute quantification of Escherichia coli proteins. In: Metabolic Engineering. 2014 ; Vol. 26. pp. 48-56.

Bibtex

@article{a23629bccceb4c1daf7868bbfab4b7c5,
title = "A targeted proteomics toolkit for high-throughput absolute quantification of Escherichia coli proteins",
abstract = "Transformation of engineered Escherichia coli into a robust microbial factory is contingent on precise control of metabolism. Yet, the throughput of omics technologies used to characterize cell components has lagged far behind our ability to engineer novel strains. To expand the utility of quantitative proteomics for metabolic engineering, we validated and optimized targeted proteomics methods for over 400 proteins from more than 20 major pathways in E. coli metabolism. Complementing these methods, we constructed a series of synthetic genes to produce concatenated peptides (QconCAT) for absolute quantification of the proteins and made them available through the Addgene plasmid repository (www.addgene.org). To facilitate high sample throughput, we developed a fast, analytical-flow chromatography method using a 5.5-min gradient (10 min total run time). Overall this toolkit provides an invaluable resource for metabolic engineering by increasing sample throughput, minimizing development time and providing peptide standards for absolute quantification of E. coli proteins.",
keywords = "Escherichia coli/metabolism, Escherichia coli Proteins/metabolism, Gene Expression Profiling/methods, High-Throughput Screening Assays/methods, Peptides/genetics, Protein Engineering/methods, Protein Interaction Mapping/methods, Proteomics/methods",
author = "Batth, {Tanveer S} and Pragya Singh and Ramakrishnan, {Vikram R} and Sousa, {Mirta M L} and Chan, {Leanne Jade G} and Tran, {Huu M} and Luning, {Eric G} and Pan, {Eva H Y} and Vuu, {Khanh M} and Keasling, {Jay D} and Adams, {Paul D} and Petzold, {Christopher J}",
note = "Copyright {\textcopyright} 2014 International Metabolic Engineering Society. All rights reserved.",
year = "2014",
doi = "10.1016/j.ymben.2014.08.004",
language = "English",
volume = "26",
pages = "48--56",
journal = "Metabolic Engineering",
issn = "1096-7176",
publisher = "Academic Press",

}

RIS

TY - JOUR

T1 - A targeted proteomics toolkit for high-throughput absolute quantification of Escherichia coli proteins

AU - Batth, Tanveer S

AU - Singh, Pragya

AU - Ramakrishnan, Vikram R

AU - Sousa, Mirta M L

AU - Chan, Leanne Jade G

AU - Tran, Huu M

AU - Luning, Eric G

AU - Pan, Eva H Y

AU - Vuu, Khanh M

AU - Keasling, Jay D

AU - Adams, Paul D

AU - Petzold, Christopher J

N1 - Copyright © 2014 International Metabolic Engineering Society. All rights reserved.

PY - 2014

Y1 - 2014

N2 - Transformation of engineered Escherichia coli into a robust microbial factory is contingent on precise control of metabolism. Yet, the throughput of omics technologies used to characterize cell components has lagged far behind our ability to engineer novel strains. To expand the utility of quantitative proteomics for metabolic engineering, we validated and optimized targeted proteomics methods for over 400 proteins from more than 20 major pathways in E. coli metabolism. Complementing these methods, we constructed a series of synthetic genes to produce concatenated peptides (QconCAT) for absolute quantification of the proteins and made them available through the Addgene plasmid repository (www.addgene.org). To facilitate high sample throughput, we developed a fast, analytical-flow chromatography method using a 5.5-min gradient (10 min total run time). Overall this toolkit provides an invaluable resource for metabolic engineering by increasing sample throughput, minimizing development time and providing peptide standards for absolute quantification of E. coli proteins.

AB - Transformation of engineered Escherichia coli into a robust microbial factory is contingent on precise control of metabolism. Yet, the throughput of omics technologies used to characterize cell components has lagged far behind our ability to engineer novel strains. To expand the utility of quantitative proteomics for metabolic engineering, we validated and optimized targeted proteomics methods for over 400 proteins from more than 20 major pathways in E. coli metabolism. Complementing these methods, we constructed a series of synthetic genes to produce concatenated peptides (QconCAT) for absolute quantification of the proteins and made them available through the Addgene plasmid repository (www.addgene.org). To facilitate high sample throughput, we developed a fast, analytical-flow chromatography method using a 5.5-min gradient (10 min total run time). Overall this toolkit provides an invaluable resource for metabolic engineering by increasing sample throughput, minimizing development time and providing peptide standards for absolute quantification of E. coli proteins.

KW - Escherichia coli/metabolism

KW - Escherichia coli Proteins/metabolism

KW - Gene Expression Profiling/methods

KW - High-Throughput Screening Assays/methods

KW - Peptides/genetics

KW - Protein Engineering/methods

KW - Protein Interaction Mapping/methods

KW - Proteomics/methods

U2 - 10.1016/j.ymben.2014.08.004

DO - 10.1016/j.ymben.2014.08.004

M3 - Journal article

C2 - 25205128

VL - 26

SP - 48

EP - 56

JO - Metabolic Engineering

JF - Metabolic Engineering

SN - 1096-7176

ER -

ID: 204046562