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

  • Organism: Escherichia coli
  • Instrument: Agilent 1290-6460
  • SpikeIn: No
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.
Created on 6/11/18, 10:03 AM
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Qcat9-Ind.sky2014-03-20 15:05:15112222662
Qcat8-Ind.sky2014-03-20 15:05:152441411271
Qcat7-Ind.sky2014-03-20 15:05:143055551652
Qcat6-Ind.sky2014-03-20 15:05:132445451343
Qcat5-Ind.sky2014-03-20 15:05:122343431291
Qcat4-Ind.sky2014-03-20 15:05:112345451341
Qcat3-Ind.sky2014-03-20 15:05:103258581731
Qcat2-Ind.sky2014-03-20 15:05:092651511501
Qcat18-Ind.sky2014-03-20 15:05:082748481422
Qcat17-Ind.sky2014-03-20 15:05:072445451351
Qcat16-Ind.sky2014-03-20 15:05:062548481441
Qcat15-Ind.sky2014-03-20 15:05:052344441321
Qcat14-Ind.sky2014-03-20 15:05:042548481633
Qcat13-Ind.sky2014-03-20 15:05:032546461373
Qcat12-Ind.sky2014-03-20 15:05:032444441361
Qcat11-Ind.sky2014-03-20 15:05:022444441392
Qcat10-Ind.sky2014-03-20 15:05:01153030903
Qcat1-Ind.sky2014-03-20 15:05:002141411221
Tyrosine-pathway-data_2014-02-28_09-56-52.sky.zip2014-02-28 09:57:431629288416
Pathways-CalCurve-Sugars-Panorama_2014-02-27_20-15-34.sky.zip2014-02-27 20:17:564096896891,36527
Limit of Detection Data_2014-02-27_16-44-26.sky.zip2014-02-27 16:47:164136136101,21915
NAD I Biosynthesis and Salvage Cycle.zip2013-05-23 04:40:2091515454
Phospholipid Anabolism.zip2013-05-22 11:04:38142626769
Tyrosine Biosynthesis.zip2013-05-21 18:07:491632329610
Tricarboxylic Acid Cycle.zip2013-05-21 18:06:001122226628
Serine Protease.zip2013-05-21 18:04:211521216327
Serine, Glycine Biosynthesis.zip2013-05-21 18:02:39488244
Purine & Pyrmidine Biosynthesis.zip2013-05-21 18:01:1834555516526
Pentose Phosphate Pathway.zip2013-05-21 17:59:51815154511
NAD Phosphorylation & Dephosphorylation.zip2013-05-21 17:56:18366182
Panthothenate-CoA Biosynthesis.zip2013-05-21 17:51:2181414423
Mixed Acid Fermentation.zip2013-05-21 17:45:161832329623
Lysine, Threonine, Methionine Biosynthesis.zip2013-05-21 17:43:1019383811417
Lipid Anabolism.zip2013-05-21 17:40:5234626218519
Leucine, Valine, Isoleucine Biosynthesis.zip2013-05-21 17:39:211528288411
Methylerythritol Phosphate (MEP) Pathway.zip2013-05-21 17:36:4571414427
Iron cluster assembly.zip2013-05-21 17:34:22713133910
Glycolysis I.zip2013-05-21 17:32:001530309025
Glycerold & glycerolphosphodiester degredation.zip2013-05-21 17:29:06488244
Glutathione Biosynthesis.zip2013-05-21 17:26:12366184
Folate Anabolism.zip2013-05-21 17:22:5481616479
Fatty Acid Biosynthesis.zip2013-05-21 17:19:02132222638
Aspartate & Asparagine Biosynthesis.zip2013-05-21 17:10:3871414427
Acetyl-CoA Biosynthesis from Pyruvate.zip2013-05-21 17:08:39366185
Acetate formation.zip2013-05-21 17:07:12488245