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PXD000095

PXD000095 is an original dataset announced via ProteomeXchange.

Dataset Summary
TitleDetermination of Phosphorylation Sites in the DivIVA Protein of Streptomyces coelicolor by Targeted LC-MS/MS
DescriptionThe filamentous bacterium Streptomyces coelicolor modulates polar growth and branching by phosphorylating the cytoskeletal protein DivIVA. Previous MALDI-TOF analysis of DivIVA showed that a large 7.2 kDa tryptic peptide was multiply phosphorylated. To aid localization of the phosphorylation sites, additional tryptic cleavage sites were introduced into DivIVA and the resulting phosphopeptides were analysed by LC-MS/MS. Phosphopeptide isomers could be separated chromatographically, but because of overlapping elution and spectrum quality, site assignment by standard software tools was ambiguous. Because fragment ions carrying the phosphate group are essential for confident localization, large numbers of spectra were collected using targeted LC-MS/MS and a special script was developed for plotting the elution of site-determining fragments from those spectra under the XIC of the parent ions. Where multiple phosphopeptide isomers were present, the elution of the site-determining y-ions perfectly coincided with the elution of the corresponding phosphopeptide isomer. This method represents a useful tool for user inspection of spectra derived from phosphopeptide isomers, and significantly increases confidence when defining phosphorylation sites. In this way, we show that DivIVA is phosphorylated in vivo on 5 sites in the C-terminal part of the protein (T304, S309, S338, S344 and S355).
HostingRepositoryPRIDE
AnnounceDate2024-10-07
AnnouncementXMLSubmission_2024-10-07_08:46:08.733.xml
DigitalObjectIdentifierhttps://dx.doi.org/10.6019/PXD000095
ReviewLevelPeer-reviewed dataset
DatasetOriginOriginal dataset
RepositorySupportSupported dataset by repository
PrimarySubmitterGerhard Saalbach
SpeciesList scientific name: Streptomyces coelicolor; NCBI TaxID: 1902;
ModificationListphosphorylated residue; monohydroxylated residue; iodoacetamide derivatized residue
InstrumentLTQ Orbitrap
Dataset History
RevisionDatetimeStatusChangeLog Entry
02012-12-11 03:53:49ID requested
12013-08-05 06:09:26announced
12014-07-28 00:26:51announced
22013-08-05 06:31:27announcedAdd reference
32024-10-07 08:46:09announced2024-10-07: Updated project metadata.
Publication List
10.6019/PXD000095;
10.1021/pr400524d;
Saalbach G, Hempel AM, Vigouroux M, Fl, รค, rdh K, Buttner MJ, Naldrett MJ, Determination of phosphorylation sites in the DivIVA cytoskeletal protein of Streptomyces coelicolor by targeted LC-MS/MS. J Proteome Res, 12(9):4187-92(2013) [pubmed]
Keyword List
submitter keyword: phosphopeptides, targeted LC-MS/MS, phosphorylation site localization, DivIVA, Orbitrap, ScaffoldPTM,Ascore
Contact List
Gerhard Saalbach
contact affiliationBiological Chemistry
contact emailgerhard.saalbach@jic.ac.uk
lab head
Gerhard Saalbach
contact affiliationBiological Chemistry
contact emailgerhard.saalbach@jic.ac.uk
dataset submitter
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Dataset FTP location
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