PXD002313

PXD002313 is an original dataset announced via ProteomeXchange.

Title	Plasmodium spp. Glyceraldehyde 3-phosphate dehydrogenase: Diverse subcellular localization and multiple post-translational modifications
Description	Glyceraldehyde 3-phosphate dehydrogenase (Gapdh), a glycolytic enzyme is known to be associated with multiple non-glycolytic functions. To examine the possibility of Plasmodium spp Gapdh having moonlighting functions, PfGapdh was cloned, overexpressed, purified to homogeneity and characterized. Polyclonal antibodies were raised in mice and used for immuno detection. Western analysis of P. yoelii whole cell extract, soluble and particulate fractions showed three major species with molecular mass ~27, 37 and 51 kDa. All three forms were equally abundant in the soluble fraction. Particulate fraction that accounted for ~10-15% of total PyGapdh did not show ~51 kDa form. Immunological evidence suggests~51 kDa form to arise by ubiquitination of native 37kDa form. For determining subcellular distribution, P. yoelii cells were subjected to differential detergent fractionation and analyzed using two-dimensional gel electrophoresis. Various subcellular fractions showed multiple species of Gapdh, indicating multiple post-translational modifications. For the determination of the nature of PTMs and residues modified, ‘Top-Down’ and ‘Bottom-Up’ approaches were used. Spots in a 2D gel that showed high intensity in western were excised and analyzed for identification of protein as well as PTMs. For more comprehensive analysis of PTM, P. yoelii whole cell extract was run on a 1D gel and bands that showed reactivity in western were cut out and analyzed by LC-MS/MS. In all, eleven residues were identified that underwent modifications. Modifications consisted of four phosphorylations, five ubiquitinations, two acetylations, two methylations and one nitrosylation. Physiological relevance of these modifications is not yet understood
HostingRepository	PRIDE
AnnounceDate	2018-01-08
AnnouncementXML	Submission_2018-01-08_00:31:37.xml
DigitalObjectIdentifier	https://dx.doi.org/10.6019/PXD002313
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	Nitin Jindal
SpeciesList	scientific name: Plasmodium yoelii yoelii; NCBI TaxID: 73239;
ModificationList	monomethylated residue; ubiquitination signature dipeptidyl lysine; phosphorylated residue; S-nitrosyl-L-cysteine; monohydroxylated residue; acetylated residue; iodoacetamide derivatized residue
Instrument	6520 Quadrupole Time-of-Flight LC/MS

Revision	Datetime	Status	ChangeLog Entry
0	2015-06-05 06:08:00	ID requested
⏵ 1	2018-01-08 00:31:38	announced

Dataset with its publication pending

curator keyword: Biological, Biomedical

submitter keyword: Plasmodium spp.

Glyceraldehyde 3-phopshate dehydrogenase

Subcellular Localization

Post-translational modification

Moonlighting functions

Mass Spectrometry

Gotam K Jarori
contact affiliation	Tata Institute of Fundamental Research, Mumbai
contact email	gkjarori@gmail.com
lab head
Nitin Jindal
contact affiliation	Tata Institute of Fundamental Research, Mumbai
contact email	nitinsjindal@gmail.com
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD002313
     1. Label: PRIDE project
     2. Name: Plasmodium spp. Glyceraldehyde 3-phosphate dehydrogenase: Diverse subcellular localization and multiple post-translational modifications