Cristea - CobB PRM

Cristea - CobB PRM
The conservation of sirtuin lipoamidase activity in bacteria as a regulator of metabolic enzyme complexes
  • Organism: Escherichia coli
  • Instrument: Q Exactive,LTQ Orbitrap Velos
  • SpikeIn: No
Abstract
Sirtuins are an essential family of nicotinamide adenine dinucleotide (NAD)-dependent enzymes, conserved from bacteria to humans. Here we study the bacterial sirtuin CobB in E. coli. To demonstrate its highly conserved enzymatic activities and substrates, we used targeted mass spectrometry to quantify CobB levels, lipoylation levels, protein interactions, and the significance of its function in various nutrient conditions.
Experiment Description
The experiments were completed using MC4100 E. coli (wild type, ΔcobB, and CobB overexpression via an IPTG-inducible promoter), and analyzed in triplicate. Total area values were exported from Skyline and normalized by the average MS1 intensity per run calculated using RawMeat. (1) CobB protein levels were measured by SRM-MS using an Orbitrap Velos. (2) Site-specific lipoylation levels of proteins ODP2 and GCSH were measured in wild type and ΔcobB cells using an Orbitrap Velos. (3) Protein interactions with His-CobB or IgG were quantified by PRM-MS using a Q-Exactive. (4) Protein levels were measured using PRM-MS with a Q-Exactive after growing cells in media with different carbon sources.
Sample Description
(1) CobB levels: Wild type and ΔcobB cells were analyzed in the file “CobB_ProteinLevels_WT_Delta_20150811.” IPTG-induced and non-induced cells were analyzed in the file “CobB_ProteinLevels_IPTG_20150805” (2) Lipoyl modification levels: “Lipoylation_SRM_MC4100_WTvDeltaCobB_20151015” (3) CobB Protein Interactions: His-CobB expressing cells were analyzed in the file “His_IP_PRM_20170414” (4) CobB and substrate protein levels in varying carbon sources: Wild type and ΔcobB cells were analyzed in the file “Carbon_Sources_PRM_MC4100_WTvDeltaCobB_20170328”
Created on 9/27/17, 2:01 PM
Clustergrammer Heatmap
 
Download
DSS Response curve_105plex_5repeats_2025-07-11_01-47-02.sky.zip2025-07-14 10:02:3296105220638600
METRIC_Eclipse_PSB_All_QCs_2025-07-10_12-02-41.sky.zip2025-07-10 09:02:4916662336
7memberCommunity_PlateB.sky.zip2025-07-02 21:47:565012122470
7memberCommunity_PlateA.sky.zip2025-07-02 21:47:565012122470
7memberCommunity_PlateD.sky.zip2025-07-02 21:47:565012122470
7memberCommunity_PlateC.sky.zip2025-07-02 21:47:565012122470
7memberCommunity_PlateB.sky.zip2025-07-02 21:47:565012122470
210723-ATI-barley-Pt14_2025-02-24_01-08-00.sky.zip2025-07-02 20:45:2110101987330
210423-ATI-barley-Pt13_2025-02-24_01-07-11.sky.zip2025-07-02 20:45:2010101940270
210416-ATI-barley-Pt12_2025-02-24_01-06-33.sky.zip2025-07-02 20:45:1810101938430
210415-ATI-barley-Pt11_2025-02-24_01-05-41.sky.zip2025-07-02 20:45:1610101940290
210302-ATI-barley-Pt10_2025-02-24_01-05-05.sky.zip2025-07-02 20:45:1510101942340
210224-ATI-barley-Pt9_2025-02-24_01-04-22.sky.zip2025-07-02 20:45:1310101940310
210217-ATI-barley-Pt8_2025-02-24_01-03-46.sky.zip2025-07-02 20:45:1210101940290
210210-ATI-barley-Pt7_2025-02-24_01-03-02.sky.zip2025-07-02 20:45:1010101946310
210203-ATI-barley-Pt6_2025-02-24_01-02-20.sky.zip2025-07-02 20:45:0910101950310
210129-ATI-barley-Pt5_2025-02-24_01-01-22.sky.zip2025-07-02 20:45:0710101958310
200731-ATI-barley-Pt4_2025-02-24_01-00-26.sky.zip2025-07-02 20:45:0610101987190
200717-ATI-barley-Pt3_2025-02-24_00-58-01.sky.zip2025-07-02 20:45:0410101987550
200709-ATI-barley-Pt2-Response_2025-02-24_00-56-48.sky.zip2025-07-02 20:45:011010199570
200709-ATI-barley-Pt2_2025-02-24_00-55-11.sky.zip2025-07-02 20:45:0110101991360
200602-ATI-barley-Pt1_2025-02-24_00-25-35.sky.zip2025-07-02 20:44:5910101993310
TPAD_CSF1A_Batch1-multiNF-DIANN.sky.zip2025-07-01 17:49:472,85327,12832,575259,532430
TPAD_CSF1A_Batch1-multiNF-DIANN-grouped.sky.zip2025-07-01 17:41:312,67226,43931,769253,119430
TPAD_CSF1B_Batch2-multiNF-DIANN.sky.zip2025-07-01 15:26:132,85327,12832,575259,532410
TPAD_CSF1B_Batch2-multiNF-DIANN-grouped.sky.zip2025-07-01 15:17:422,67226,43931,769253,119410
TPAD_CSF2A_Batch3-multiNF-DIANN.sky.zip2025-07-01 14:13:302,85327,12832,575259,532430
TPAD_CSF2A_Batch3-multiNF-DIANN-grouped.sky.zip2025-07-01 14:04:582,67226,43931,769253,119430
TPAD_CSF2B_Batch4-multiNF-DIANN.sky.zip2025-07-01 11:21:372,85327,12832,575259,532430
TPAD_CSF2B_Batch4-multiNF-DIANN-grouped.sky.zip2025-07-01 11:09:482,67226,43931,769253,119430
TPAD_CSF3A_Batch5-multiNF-DIANN.sky.zip2025-06-30 15:31:332,85327,12832,575259,532430
TPAD_CSF3A_Batch5-multiNF-DIANN-grouped.sky.zip2025-06-30 15:23:252,67226,43931,769253,119430
TPAD_CSF3B_Batch6-multiNF-DIANN.sky.zip2025-06-30 14:43:542,85327,12832,575259,532410
TPAD_CSF3B_Batch6-multiNF-DIANN-grouped.sky.zip2025-06-30 14:36:142,67226,43931,769253,119410
TPAD_CSF4_Batch7-multiNF-DIANN.sky.zip2025-06-30 13:55:172,85327,12832,575259,532420
TPAD_CSF4_Batch7-multiNF-DIANN-grouped.sky.zip2025-06-30 13:46:482,67226,43931,769253,119420
TPAD_CSF5_Batch8-multiNF-DIANN.sky.zip2025-06-30 13:02:062,85327,12832,575259,532540
TPAD_CSF5_Batch8-multiNF-DIANN-grouped.sky.zip2025-06-30 11:52:172,67226,43931,769253,119540
Exp_CSF_GPF_A_2024-05-06_21-16-47.sky.zip2025-06-25 18:12:201,8297,3787,37855,04010
2024-05 CSF LIT GPF-MSFragger-3.sky.zip2025-06-25 18:12:201,0825,0956,01940,40160
240207_gpf_2024-06-04_10-24-18.sky.zip2025-06-25 18:12:201,2886,98410,593126,08610
230305_ev2_prm_final_inj1_2024-06-04_07-32-10.sky.zip2025-06-25 18:12:207222,0892,08914,449420
230124_p2_neo_30min_3500targets_opt_trans_pepleveldilution_2024-06-04_06-58-16.sky.zip2025-06-25 18:12:201,0273,5013,50111,420410
CSF_neurod105_assay_individuals_manual_2024-06-03_15-26-36.sky.zip2025-06-25 18:12:201029029026,345370
Exp_CSF_MMCC_quant_all_adjBound_opttrans_nochick_2024-06-03_14-53-02.sky.zip2025-06-25 18:12:201,2058,3748,37424,934270
OT_GPF_PRM_survey_MMCC_boundaries_opttrans_nochick_2024-06-03_14-36-08.sky.zip2025-06-25 18:12:201,2801,9711,9715,801270
LIT_GPF_survey_newAlign_MMCC_boundaries_opttrans_nochick_2024-06-02_15-26-11.sky.zip2025-06-25 18:12:207982,0352,0355,998270
Aducanumab_Selectivity_2025-06-19_12-58-48.sky.zip2025-06-19 09:59:06448512364
hela_isolation_window_concentration_2000ng.sky.zip2025-06-02 18:36:1910,365188,947223,7653,101,148180
hela_isolation_window_experiment_200ng.sky.zip2025-06-02 18:36:1910,365188,947223,7653,101,148180
hela_isolation_window_experiment_500ng.sky.zip2025-06-02 18:36:1910,364188,947223,7653,101,148180
hela_isolation_window_experiment_1000ng.sky.zip2025-06-02 18:36:1910,365188,947223,7653,101,148180
2025-0405-MMCC-EV-Astral-noimpute_2025-05-29_11-52-05.sky.zip2025-06-02 18:36:194,90737,54742,331335,891420
2025-0405-MMCC-EV-Actis-noimpute_2025-05-29_11-46-45.sky.zip2025-06-02 18:36:195,18541,97847,810380,072420
SMTG-low-value-imputed_2025-05-01_14-22-15.sky.zip2025-06-01 21:38:1710,383187,220217,1092,214,248780
SMTG-MissForest-imputed_2025-05-01_11-41-41.sky.zip2025-06-01 20:04:5710,383187,220217,1092,214,248780
SMTG-nettle-imputed_2025-04-30_12-25-18.sky.zip2025-06-01 18:16:1410,639215,139251,4792,483,959780
SMTG-unimputed_2025-04-30_08-55-40.sky.zip2025-06-01 13:32:5610,639215,139251,4792,483,959780
TEI-REX-unimputed_2025-04-29_15-46-15.sky.zip2025-06-01 11:49:381,30749,85259,007585,987960
TEI-REX-nettle-imputed_2025-04-29_13-36-50.sky.zip2025-06-01 11:15:251,30749,85259,007585,987960
MagNet-holdout-nettle-imputed_2025-05-01_08-50-25.sky.zip2025-06-01 09:40:175,29914,11015,07460,288420
hela_isolation_window_concentration_2000ng.sky.zip2025-05-30 15:25:4610,365188,947223,7653,101,148180
ecoli_large_replicates_loaded_refined_2024-05-30_14-02-04.sky.zip2025-05-30 12:00:183669899894,94080
ecoli_large_replicates_loaded_2024-05-30_14-00-59.sky.zip2025-05-30 12:00:184611,3021,3027,52480
ecoli_large_replicates_2024-05-30_14-00-05.sky.zip2025-05-30 12:00:184611,3021,3027,52400
ecoli_subset_replicates_refined_cv_2024-05-30_13-58-18.sky.zip2025-05-30 12:00:187592,2662,26612,80720
ecoli_subset_replicates_refined_2024-05-30_13-57-25.sky.zip2025-05-30 12:00:187592,4372,43713,60220
ecoli_subset_replicates_2024-05-30_13-56-16.sky.zip2025-05-30 12:00:187592,4372,44130,85720
gpf_results_importer_2024-05-30_13-54-14.sky.zip2025-05-30 12:00:181,1925,3967,42785,67310
gpf_results_manual_2024-05-30_13-52-23.sky.zip2025-05-30 12:00:181,1925,3967,42785,67310
pq500_100spd_plasma_final_lightheavy_replicates_2024-05-30_13-50-10.sky.zip2025-05-30 11:58:415798181,62213,699100
pq500_60spd_plasma_final_lightheavy_replicates_2024-05-30_13-48-26.sky.zip2025-05-30 11:58:415798181,62213,876100
pq500_100spd_plasma_final_replicates_2024-05-30_13-47-38.sky.zip2025-05-30 11:58:415798188187,081100
pq500_60spd_plasma_final_replicates_2024-05-30_13-46-43.sky.zip2025-05-30 11:58:415798188186,977100
pq500_100spd_plasma_multireplicate_results_refined_2024-05-30_13-46-07.sky.zip2025-05-30 11:58:415798188187,08110
pq500_60spd_plasma_multireplicate_results_refined_2024-05-30_13-45-40.sky.zip2025-05-30 11:58:415798188186,97710
pq500_100spd_plasma_multireplicate_results_2024-05-30_13-45-07.sky.zip2025-05-30 11:58:415798188187,65120
pq500_60spd_plasma_multireplicate_results_2024-05-30_13-44-21.sky.zip2025-05-30 11:58:415798188187,60120
pq500_100spd_neat_multireplicate_results_refined_2024-05-30_13-43-42.sky.zip2025-05-30 11:58:415798188187,68910
pq500_60spd_neat_multireplicate_results_refined_2024-05-30_13-43-05.sky.zip2025-05-30 11:58:415798188187,60110
pq500_100spd_neat_multireplicate_results_2024-05-30_13-42-26.sky.zip2025-05-30 11:58:4157981881811,08810
pq500_60spd_neat_multireplicate_results_2024-05-30_13-41-26.sky.zip2025-05-30 11:58:4157981881811,08810
pq500_60spd_neat_multireplicate_2024-05-30_13-40-53.sky.zip2025-05-30 11:58:4157981881811,08800
hela_isolation_window_experiment_200ng.sky.zip2025-05-30 01:29:3910,365188,947223,7653,101,148180
hela_isolation_window_experiment_500ng.sky.zip2025-05-30 00:53:5310,364188,947223,7653,101,148180
hela_isolation_window_experiment_1000ng.sky.zip2025-05-29 23:15:1110,365188,947223,7653,101,148180
2025-0405-MMCC-EV-Astral-noimpute_2025-05-29_11-52-05.sky.zip2025-05-29 12:13:324,90737,54742,331335,891420
2025-0405-MMCC-EV-Actis-noimpute_2025-05-29_11-46-45.sky.zip2025-05-29 12:03:295,18541,97847,810380,072420
20240709_OBIPhA.sky.zip2025-05-16 09:25:5133337235120
Swaney-IPMS-Data-DIANN_2025-05-15_11-53-43.sky.zip2025-05-15 12:02:265,18545,78152,836422,661150
Vial Comparison Post 01152025 v1 (not daily)_2025-04-29_15-52-29.sky.zip2025-04-30 21:22:23102424190
Vial Comparison Pre v2_2025-04-29_15-49-25.sky.zip2025-04-30 21:22:23102424230
ATI_Samples_2023_2025-04-26_13-39-03.sky.zip2025-04-28 10:30:14132448280420
ATI_Response_2023_2025-04-26_13-25-06.sky.zip2025-04-28 10:30:14132448280180
ATI_Samples_2022_2025-02-20_23-31-07.sky.zip2025-04-28 10:30:10122346268480
ATI_Samples_2021_2025-02-20_23-21-52.sky.zip2025-04-28 10:30:08122346268300
2023_ATIsamples_circe_2025-02-20_13-24-11.sky.zip2025-04-25 14:58:17132243250960
2022_ATIsamples_circe_2025-02-20_13-17-06.sky.zip2025-04-25 14:58:09122243250970
2021_ATIsamples_circe_2025-02-20_13-06-53.sky.zip2025-04-25 14:58:02122243250940
deamidation_localization_gS_7-18_2025-04-24_20-52-46.sky.zip2025-04-25 12:05:181441810