Project PXD003556

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Biomedical Dataset
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Summary

Title

The proteomes of plasmodium falciparum mature male and female gametocytes

Description

Gametocytes are obligate for the transmission of malaria parasite to mosquito. This study reports the first comprehensive proteomes of accurately separated mature male and female Plasmodium falciparum gametocytes using a transgenic line. the proteomes revealed significant differences that are consistent with the divergent functions of the two sexes. While the male proteome is enriched in proteins associated with flagella and genome replication, the female proteome is more abundant in proteins involved in metabolism, translation and organellar functions.

Sample Processing Protocol

The separated parasites were lysed in SDT-lysis buffer (4% SDS, 100 mM Tris/HCl pH 7.6, 0.1 M DTT) for complete dissolution and separated in a 10% Bis-Tris SDS-PAGE gel. Ten slices were excised in-gel digestion was done as described (Lasonder et al., 2002). The digests were analyzed by nano LC/MS/MS with a Waters NanoAcquity HPLC system interfaced to a Q Exactive™ Hybrid Quadrupole-Orbitrap Mass Spectrometer (Thermo Scientific). Peptides were loaded on a trapping column and eluted over a 75 μm analytical column at 350 nL/min. MS and MS/MS were performed at 70,000 FWHM and 17,500 FWHM resolution, respectively. The fifteen most abundant ions were selected for MS/MS.

Data Processing Protocol

The LC/MS/MS data were searched against Uniprot P. falciparum protein database (v01/2014) with reversed entries in order to assess false discovery rate (FDR). Data were filtered at 0.7-1% protein and 0.1-0.2% peptide FDR. Search parameters included trypsin digestion (C-terminal K and R cleavage), full cleavage with 2 missed sites, fragment ion mass tolerance with tolerance of 0.02 Da, peptide mass tolerance of 10 ppm. Mascot DAT files were parsed into the Scaffold software for validation and filtering to create a non-redundant list per sample.

Contact

JUN MIAO, Pennsylvania State University
Liwang Cui, Department of Entomology, Pennsylvania State University, USA ( lab head )

Submission Date

01/02/2016

Publication Date

30/01/2017

Publication

    Miao J, Chen Z, Wang Z, Shrestha S, Li X, Li R, Cui L. Sex-Specific Biology of the Human Malaria Parasite Revealed from the Proteomes of Mature Male and Female Gametocytes. Mol Cell Proteomics. 2017 Jan 26. pii: mcp.M116.061804 PubMed: 28126901

Assay

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# Accession Title Proteins Peptides Unique Peptides Spectra Identified Spectra View in Reactome
1 61600 no assay title provided (mzIdentML) 426 3094 1579 1782 1782
2 61602 no assay title provided (mzIdentML) 407 2934 1958 2231 2231
3 61601 no assay title provided (mzIdentML) 482 3094 2240 2402 2402
4 61604 no assay title provided (mzIdentML) 513 3944 2780 3209 3209
5 61640 no assay title provided (mzIdentML) 528 4208 2563 3164 3164
6 61603 no assay title provided (mzIdentML) 540 5460 3095 4049 4049
7 61641 no assay title provided (mzIdentML) 674 4183 2779 3375 3375
8 61642 no assay title provided (mzIdentML) 640 3293 2194 2631 2631
9 61643 no assay title provided (mzIdentML) 553 2215 1429 1629 1629
10 61644 no assay title provided (mzIdentML) 384 1577 794 948 948