Summary of Study ST000453

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR000350. The data can be accessed directly via it's Project DOI: 10.21228/M8T30C This work is supported by NIH grant, U2C- DK119886.

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Study IDST000453
Study TitleMetabolic Adaptation of Staphylococcus aureus to Host Immunity
Study TypeMetabolomics Analysis of Methicillin-Resistant Staphylococcus aureus (MRSA) to host immunity.
Study SummaryThis project is intended to study the metabolic adaptation of Methicillin-Resistant Staphylococcus aureus (MRSA) to host immunity. Because of the nature of the samples RTI RCMRC worked with Dr. Anthony R. Richardson so that the samples would be extracted at the University of North Carolina at Chapel Hill under the condition that were optimized by RTI RCMRC for broad spectrum metabolomics analysis.
Institute
University of North Carolina
DepartmentRCMRC
LaboratorySumner Lab
Last NameSumner
First NameSusan
AddressEastern Regional Comprehensive Metabolomics Resource Core, UNC Nutrition Research Institute, 500 Laureate Way, Kannapolis, NC, 28081
Emailsusan_sumner @unc.edu
Phone704-250-5066
Submit Date2017-09-10
Num Groups29
Total Subjects29
Raw Data AvailableYes
Raw Data File Type(s)wiff
Analysis Type DetailLC-MS
Release Date2017-10-03
Release Version1
Susan Sumner Susan Sumner
https://dx.doi.org/10.21228/M8T30C
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Project:

Project ID:PR000350
Project DOI:doi: 10.21228/M8T30C
Project Title:Metabolic Adaptation of Staphylococcus aureus to Host Immunity
Project Summary:Staphylococcus aureus infections are difficult to treat given the drug-resistant nature of this bacterium. Methicillin-Resistant Staphylococcus aureus (MRSA) is considered the single most deadly bacterial infection in the United States. The bacterium resists nearly every aspect of innate immunity, including the cytotoxic effects of nitric oxide (NO?) which is an innate immune effector produced by phagocytes in response to infection. NO? interferes with various metabolic pathways in pathogens by reacting with redox centers of multiple enzymes. We have demonstrated that S. aureus NO?-resistance is essential for full virulence. We have also shown that S. aureus NO-resistance is predicated on the abundance of glucose, which may explain exacerbated S. aureus disease outcomes associated with diabetic patients having poorly controlled blood sugar. Here we propose to employ metabolomics to probe the nature of the S. aureus NO-resistant metabolic state. We will thereby bridge a fundamental gap in our knowledge of S. aureus pathogenesis by defining metabolic pathways essential to cause disease in immunocompetent hosts. These key metabolic pathways represent important therapeutic targets for future research. Define the requirement for glycolytic carbon/energy sources for the S. aureus NO-RMS: While S. aureus can thrive on a variety of gluconeogenic carbon sources (e.g. amino acids, glycerol, lactate etc.), none of these are capable of supporting NO?-resistance. We will compare changes in intracellular metabolites in cells growing on gluconeogenic carbon sources following NO?-exposure to elucidate what features of gluconeogenesis are incompatible with NO?-resistance. The absolute requirement for glycolytic carbon sources for NO-resistance is interesting given the disproportionately poor disease outcomes of MRSA infections in diabetic patients with elevated blood glucose.
Institute:University of North Carolina at Chapel Hill
Department:Department of Microbiology and Immnunology, UNC-CH
Last Name:Richardson
First Name:Anthony
Address:125 Mason Farm Rd, Marsico Hall 6209, Chapel Hill, NC 27599
Email:anthony_richardson@med.unc.edu
Phone:919-843-3654

Subject:

Subject ID:SU000474
Subject Type:Bacterial Cells
Subject Species:Staphylococcus aureus
Taxonomy ID:1280
Species Group:Microorganism

Factors:

Subject type: Bacterial Cells; Subject species: Staphylococcus aureus (Factor headings shown in green)

mb_sample_id local_sample_id Cell Additive 1 Additive 2 Environment NO status Time
SA022900Total_Pool_02- - - - - -
SA022901Total_Pool_01- - - - - -
SA022902Total_Pool_03- - - - - -
SA022903Total_Pool_04- - - - - -
SA022904Total_Pool_05- - - - - -
SA022908CE_mut_G_NO_1_5_27_Cmutant Glucose none undefined NO 1.5 h
SA022909CE_mut_G_NO_1_5_27_Amutant Glucose none undefined NO 1.5 h
SA022910CE_mut_G_NO_1_5_27_Bmutant Glucose none undefined NO 1.5 h
SA022905CE_mut_G_NO_1_26_Bmutant Glucose none undefined NO 1 h
SA022906CE_mut_G_NO_1_26_Cmutant Glucose none undefined NO 1 h
SA022907CE_mut_G_NO_1_26_Amutant Glucose none undefined NO 1 h
SA022911CE_mut_G_NO_3_28_Cmutant Glucose none undefined NO 5 h
SA022912CE_mut_G_NO_3_28_Bmutant Glucose none undefined NO 5 h
SA022913CE_mut_G_NO_3_28_Amutant Glucose none undefined NO 5 h
SA022914CE_mut_G_NO_4_29_Amutant Glucose none undefined NO 6 h
SA022915CE_mut_G_NO_4_29_Bmutant Glucose none undefined NO 6 h
SA022916CE_mut_G_NO_4_29_Cmutant Glucose none undefined NO 6 h
SA022917CE_AA_A_3_Anormal AA none Aerobic not specified not specified
SA022918CE_AA_A_3_Cnormal AA none Aerobic not specified not specified
SA022919CE_AA_A_3_Bnormal AA none Aerobic not specified not specified
SA022920CE_AA_NO_0_25_18_Cnormal AA none undefined NO 0.25 h
SA022921CE_AA_NO_0_25_18_Anormal AA none undefined NO 0.25 h
SA022922CE_AA_NO_0_25_18_Bnormal AA none undefined NO 0.25 h
SA022923CE_AA_NO_0_5_19_Cnormal AA none undefined NO 0.5 h
SA022924CE_AA_NO_0_5_19_Anormal AA none undefined NO 0.5 h
SA022925CE_AA_NO_0_5_19_Bnormal AA none undefined NO 0.5 h
SA022926CE_AA_NO_1_5_21_Anormal AA none undefined NO 1.5 h
SA022927CE_AA_NO_1_5_21_Cnormal AA none undefined NO 1.5 h
SA022928CE_AA_NO_1_5_21_Bnormal AA none undefined NO 1.5 h
SA022932CE_AA_NO_2_5_23_Anormal AA none undefined NO 2.5 h
SA022933CE_AA_NO_2_5_23_Bnormal AA none undefined NO 2.5 h
SA022934CE_AA_NO_2_5_23_Cnormal AA none undefined NO 2.5 h
SA022929CE_AA_NO_2_17_Bnormal AA none undefined NO 2 h
SA022930CE_AA_NO_2_17_Anormal AA none undefined NO 2 h
SA022931CE_AA_NO_2_17_Cnormal AA none undefined NO 2 h
SA022935CE_AA_NO_3_24_Anormal AA none undefined NO 3 h
SA022936CE_AA_NO_3_24_Cnormal AA none undefined NO 3 h
SA022937CE_AA_NO_1_20_Bnormal AA none undefined NO 3 h
SA022938CE_AA_NO_3_24_Bnormal AA none undefined NO 3 h
SA022939CE_AA_NO_1_20_Cnormal AA none undefined NO 3 h
SA022940CE_AA_NO_1_20_Anormal AA none undefined NO 3 h
SA022941CE_AA_NO_4_25_Anormal AA none undefined NO 4 h
SA022942CE_AA_NO_4_25_Cnormal AA none undefined NO 4 h
SA022943CE_AA_NO_4_25_Bnormal AA none undefined NO 4 h
SA022944CE_AA_NO_2_22_Anormal AA none undefined NO 4 h
SA022945CE_AA_NO_2_22_Bnormal AA none undefined NO 4 h
SA022946CE_AA_NO_2_22_Cnormal AA none undefined NO 4 h
SA022947CE_AA_P_A_4_Anormal AA pyruvate Aerobic not specified not specified
SA022948CE_AA_P_A_4_Cnormal AA pyruvate Aerobic not specified not specified
SA022949CE_AA_P_A_4_Bnormal AA pyruvate Aerobic not specified not specified
SA022950CE_G_A_1_Anormal Glucose none Aerobic not specified not specified
SA022951CE_G_A_1_Cnormal Glucose none Aerobic not specified not specified
SA022952CE_G_A_1_Bnormal Glucose none Aerobic not specified not specified
SA022953CE_G_AN_NO2_8_Anormal Glucose none Anaerobic NO2 not specified
SA022954CE_G_AN_NO2_8_Bnormal Glucose none Anaerobic NO2 not specified
SA022955CE_G_AN_NO2_8_Cnormal Glucose none Anaerobic NO2 not specified
SA022956CE_G_AN_NO3_7_Cnormal Glucose none Anaerobic NO3 not specified
SA022957CE_G_AN_NO3_7_Bnormal Glucose none Anaerobic NO3 not specified
SA022958CE_G_AN_NO3_7_Anormal Glucose none Anaerobic NO3 not specified
SA022959CE_G_AN_5_Cnormal Glucose none Anaerobic not specified not specified
SA022960CE_G_AN_5_Anormal Glucose none Anaerobic not specified not specified
SA022961CE_G_AN_5_Bnormal Glucose none Anaerobic not specified not specified
SA022962CE_G_HY_6_Cnormal Glucose none Hypoxic not specified not specified
SA022963CE_G_HY_6_Bnormal Glucose none Hypoxic not specified not specified
SA022964CE_G_HY_6_Anormal Glucose none Hypoxic not specified not specified
SA022965CE_G_NO_0_25_9_Anormal Glucose none undefined NO 0.25 h
SA022966CE_G_NO_0_25_9_Cnormal Glucose none undefined NO 0.25 h
SA022967CE_G_NO_0_25_9_Bnormal Glucose none undefined NO 0.25 h
SA022968CE_G_NO_0_5_10_Anormal Glucose none undefined NO 0.5 h
SA022969CE_G_NO_0_5_10_Bnormal Glucose none undefined NO 0.5 h
SA022970CE_G_NO_0_5_10_Cnormal Glucose none undefined NO 0.5 h
SA022974CE_G_NO_1_5_12_Cnormal Glucose none undefined NO 1.5 h
SA022975CE_G_NO_1_5_12_Anormal Glucose none undefined NO 1.5 h
SA022976CE_G_NO_1_5_12_Bnormal Glucose none undefined NO 1.5 h
SA022971CE_G_NO_1_11_Anormal Glucose none undefined NO 1 h
SA022972CE_G_NO_1_11_Cnormal Glucose none undefined NO 1 h
SA022973CE_G_NO_1_11_Bnormal Glucose none undefined NO 1 h
SA022980CE_G_NO_2_5_14_Cnormal Glucose none undefined NO 2.5 h
SA022981CE_G_NO_2_5_14_Bnormal Glucose none undefined NO 2.5 h
SA022982CE_G_NO_2_5_14_Anormal Glucose none undefined NO 2.5 h
SA022977CE_G_NO_2_13_Anormal Glucose none undefined NO 2 h
SA022978CE_G_NO_2_13_Bnormal Glucose none undefined NO 2 h
SA022979CE_G_NO_2_13_Cnormal Glucose none undefined NO 2 h
SA022983CE_G_NO_3_15_Bnormal Glucose none undefined NO 3 h
SA022984CE_G_NO_3_15_Cnormal Glucose none undefined NO 3 h
SA022985CE_G_NO_3_15_Anormal Glucose none undefined NO 3 h
SA022986CE_G_NO_5_16_Bnormal Glucose none undefined NO 5 h
SA022987CE_G_NO_5_16_Cnormal Glucose none undefined NO 5 h
SA022988CE_G_NO_5_16_Anormal Glucose none undefined NO 5 h
SA022989CE_G_P_A_2_Cnormal Glucose pyruvate Aerobic not specified not specified
SA022990CE_G_P_A_2_Anormal Glucose pyruvate Aerobic not specified not specified
SA022991CE_G_P_A_2_Bnormal Glucose pyruvate Aerobic not specified not specified
Showing results 1 to 92 of 92

Collection:

Collection ID:CO000468
Collection Summary:Cultures of S. aureus strain COL
Sample Type:Bacterial cells

Treatment:

Treatment ID:TR000488
Treatment Summary:Glucose NO 5 h AA NO 3 h Glucose Anaer Glucose NO 2 h Glucose Hypoxic Glucose Aerobic a.a. + Pyr Aer Glucose NO .25 h Glucose Anaerobic NO3 AA NO 1.5 h Glucose NO 3 h AA NO 2 h mut Gluc NO 1.5 h Glucose Anaerobic NO2 mut Gluc NO 3 h AA NO .5 h mut Gluc NO 4 h AA NO .25 h Glucose NO 1 h mut Gluc NO 1 h AA NO 4 h AA NO 2.5 h Glucose NO 2.5 h AA NO 1 h Glucose NO 1.5 h AA NO 2 h Glucose NO .5 h a.a. Aer Glucose + Pyruvate Aerobic

Sample Preparation:

Sampleprep ID:SP000481
Sampleprep Summary:Methylillin-resistant S. aureus (MRSA) bacterial culture extracts re-suspended in water were provided to RTI RCMRC for analysis. These extracts were stored at -80 °C upon arrival. For processing, samples were dried overnight on a lyophilizer. The residue was then reconstituted with 150 µL of an ice-cold solution of reconstitution buffer (0.005 mg/mL Tryptophan-d5 in 95:5 Water:Methanol, v/v) and mixed on a multiple tube vortexer for 2 min at 5,000 rpm. Then, the samples were centrifuged at room temperature for 4 min at 16,000 rcf, and 100 µL of the supernatants were transferred to autosampler vials for data acquisition. Whole study pooled QC samples were created by combining 40 µL aliquot from each of the reconstituted samples into a 2 mL LoBind eppendorf tube. The QC pooled sample was then vortexed for 15 sec and 100 µL of the total study pooled QC samples were transferred to five pre-labeled autosampler vials.

Combined analysis:

Analysis ID AN000709 AN000710
Analysis type MS MS
Chromatography type Reversed phase Reversed phase
Chromatography system Waters Acquity Waters Acquity
Column Waters Acquity HSS T3 (100 x 2.1mm,1.8um) Waters Acquity HSS T3 (100 x 2.1mm,1.8um)
MS Type ESI ESI
MS instrument type QTOF QTOF
MS instrument name Waters Synapt G2 Si QTOF Waters Synapt G2 Si QTOF
Ion Mode POSITIVE NEGATIVE
Units counts counts

Chromatography:

Chromatography ID:CH000513
Instrument Name:Waters Acquity
Column Name:Waters Acquity HSS T3 (100 x 2.1mm,1.8um)
Column Pressure:6,000-10,000 psi
Column Temperature:50 °C
Flow Rate:0.4 mL/min
Injection Temperature:8 °C
Solvent A:100% water; 0.1% formic acid
Solvent B:100% methanol; 0.1% formic acid
Analytical Time:22 min
Randomization Order:Yes
Chromatography Type:Reversed phase

MS:

MS ID:MS000631
Analysis ID:AN000709
Instrument Name:Waters Synapt G2 Si QTOF
Instrument Type:QTOF
MS Type:ESI
Ion Mode:POSITIVE
  
MS ID:MS000632
Analysis ID:AN000710
Instrument Name:Waters Synapt G2 Si QTOF
Instrument Type:QTOF
MS Type:ESI
Ion Mode:NEGATIVE
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