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Validating Missing Proteins in Human Sperm Cells by Targeted Mass-Spectrometry- and Antibody-based Methods
Univ Strasbourg, Lab Spectrometrie Masse BioOrgan LSMBO, IPHC, CNRS,UMR7178, 25 Rue Becquerel, F-67087 Strasbourg, France..
SIB, CALIPHO Grp, CMU, Rue Michel Servet 1, CH-1211 Geneva 4, Switzerland..
Univ Strasbourg, Lab Spectrometrie Masse BioOrgan LSMBO, IPHC, CNRS,UMR7178, 25 Rue Becquerel, F-67087 Strasbourg, France..
Univ Rennes 1, UMS BioSit, Core Facil H2P2, F-35040 Rennes, France..
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2017 (English)In: Journal of Proteome Research, ISSN 1535-3893, E-ISSN 1535-3907, Vol. 16, no 12, p. 4340-4351Article in journal (Refereed) Published
Abstract [en]

The present study is a contribution to the "neXt50 challenge", a coordinated effort across C-HPP teams to identify the SO most tractable missing proteins (MPs) on each chromosome. We report the targeted search of 38 theoretically detectable MPs from chromosomes 2 and 14 in Triton X-100 soluble and insoluble sperm fractions from a total of 15 healthy donors. A targeted mass spectrometry-based strategy consisting of the development of LC-PRM assays (with heavy labeled synthetic peptides) targeting 92 proteotypic peptides of the 38 selected MPs was used. Out of the 38 selected MPs, 12 were identified with two or more peptides and 3 with one peptide after extensive SDS-PAGE fractionation of the two samples and with overall low-intensity signals. The PRM data are available via ProteomeXchange in PASSEL (PASS01013). Further validation by immunohistochemistry on human testes sections and cytochemistry on sperm smears was performed for eight MPs with antibodies available from the Human Protein Atlas. Deep analysis of human sperm still allows the validation of MPs and therefore contributes to the C-HPP worldwide effort. We anticipate that our results will be of interest to the reproductive biology community because an in-depth analysis of these MPs may identify potential new candidates in the context of human idiopathic infertilities.

Place, publisher, year, edition, pages
2017. Vol. 16, no 12, p. 4340-4351
Keywords [en]
human proteome project, spermatozoon, missing proteins, parallel reaction monitoring, targeted proteomics, immunohistochemistry, immunocytochemistry, bioinformatics, data mining
National Category
Biochemistry and Molecular Biology
Identifiers
URN: urn:nbn:se:uu:diva-344202DOI: 10.1021/acs.jproteome.7b00374ISI: 000417228900010PubMedID: 28891297OAI: oai:DiVA.org:uu-344202DiVA, id: diva2:1187750
Available from: 2018-03-05 Created: 2018-03-05 Last updated: 2018-03-08Bibliographically approved

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Lindskog, Cecilia

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