PXD009614 is an
original dataset announced via ProteomeXchange.
Dataset Summary
Title | Quantitative analysis of the exoproteome from a Staphylococcus aureus LAC sarA mutant |
Description | The staphylococcal accessory regulator A (sarA) impacts the extracellular accumulation of Staphylococcus aureus virulence factors at the level of intracellular production and extracellular protease-mediated degradation. To assess the relative impact of these two functions, we previously used a proteomics approach that measures protein abundance as a function of all proteoforms to demonstrate that mutation of sarA results in increased levels of extracellular proteases and assess the impact of this on the accumulation of S. aureus exoproteins1. While this approach confirmed that protease-mediated degradation has a significant impact on the S. aureus exoproteome, it was potentially limited in that it did not take into account the possibility that large, stable proteolytic products from a given protein could result in false negatives when quantified by total proteoforms. Here, we present an expanded proteomics approach that utilizes a dual quantitative method for measuring abundance at both the total proteoform and full-length exoprotein levels. Specifically, proteins present in conditioned medium from overnight, stationary phase cultures of the USA300 strain LAC, an isogenic sarA mutant, and a sarA mutant unable to produce any of the known extracellular proteases (sarA/protease) were resolved using one-dimensional gel electrophoresis. Using methods that focus on total proteoforms vs. methods that focus specifically on full-length proteins, quantitative proteomic comparisons of sarA vs sarA/protease mutants identified proteins that were degraded in a protease dependent manner owing to mutation of sarA, while comparisons of a sarA/protease mutant vs the LAC parent strain identified proteins in which abundance was altered in a sarA mutant in a protease-independent manner. Furthermore, the proteins uniquely identified by the full-length data analysis approach eliminated false negatives observed in the total proteoform analysis. This approach provided for a more comprehensive and robust analysis of the impact of mutating sarA and protease-mediated degradation on the S. aureus exoproteome. |
HostingRepository | PRIDE |
AnnounceDate | 2018-11-21 |
AnnouncementXML | Submission_2018-11-21_06:01:52.xml |
DigitalObjectIdentifier | https://dx.doi.org/10.6019/PXD009614 |
ReviewLevel | Peer-reviewed dataset |
DatasetOrigin | Original dataset |
RepositorySupport | Supported dataset by repository |
PrimarySubmitter | Stephanie Byrum |
SpeciesList | scientific name: Staphylococcus aureus; NCBI TaxID: 1280; |
ModificationList | No PTMs are included in the dataset |
Instrument | LTQ Orbitrap |
Dataset History
Revision | Datetime | Status | ChangeLog Entry |
0 | 2018-04-26 03:15:56 | ID requested | |
⏵ 1 | 2018-11-21 06:01:53 | announced | |
Publication List
Byrum SD, Loughran AJ, Beenken KE, Orr LM, Storey AJ, Mackintosh SG, Edmondson RD, Tackett AJ, Smeltzer MS, Label-Free Proteomic Approach to Characterize Protease-Dependent and -Independent Effects of sarA Inactivation on the Staphylococcus aureus Exoproteome. J Proteome Res, 17(10):3384-3395(2018) [pubmed] |
Keyword List
curator keyword: Biological, Biomedical |
submitter keyword: protease, proteolysis, Staphylococcus aureus, sarA, GelCMS |
Contact List
Alan J. Tackett |
contact affiliation | Department of Biochemistry and Molecular Biology, University of Arkansas for Medical Sciences, 4301 West Markham Street, Little Rock, AR 72205, USA Arkansas Children's Research Institute, 13 Children's Way, Little Rock, AR 72205, USA |
contact email | ajtackett@uams.edu |
lab head | |
Stephanie Byrum |
contact affiliation | UAMS |
contact email | sbyrum@uams.edu |
dataset submitter | |
Full Dataset Link List
Dataset FTP location
NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2018/11/PXD009614 |
PRIDE project URI |
Repository Record List
[ + ]
[ - ]
- PRIDE
- PXD009614
- Label: PRIDE project
- Name: Quantitative analysis of the exoproteome from a Staphylococcus aureus LAC sarA mutant