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  • Carapito, ChristineUniv Strasbourg, Lab Spectrometrie Masse BioOrgan LSMBO, IPHC, CNRS,UMR7178, 25 Rue Becquerel, F-67087 Strasbourg, France. (author)

Validating Missing Proteins in Human Sperm Cells by Targeted Mass-Spectrometry- and Antibody-based Methods

  • Article/chapterEnglish2017

Publisher, publication year, extent ...

  • 2017-09-26
  • American Chemical Society (ACS),2017
  • printrdacarrier

Numbers

  • LIBRIS-ID:oai:DiVA.org:uu-344202
  • https://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-344202URI
  • https://doi.org/10.1021/acs.jproteome.7b00374DOI

Supplementary language notes

  • Language:English
  • Summary in:English

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  • Subject category:ref swepub-contenttype
  • Subject category:art swepub-publicationtype

Notes

  • 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.

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  • Duek, PaulaSIB, CALIPHO Grp, CMU, Rue Michel Servet 1, CH-1211 Geneva 4, Switzerland. (author)
  • Macron, CharlotteUniv Strasbourg, Lab Spectrometrie Masse BioOrgan LSMBO, IPHC, CNRS,UMR7178, 25 Rue Becquerel, F-67087 Strasbourg, France. (author)
  • Seffals, MarineUniv Rennes 1, UMS BioSit, Core Facil H2P2, F-35040 Rennes, France. (author)
  • Rondel, KarineIrset, INSERM, U1085, Protim, Campus Beaulieu, F-35042 Rennes, France. (author)
  • Delalande, FrancoisUniv Strasbourg, Lab Spectrometrie Masse BioOrgan LSMBO, IPHC, CNRS,UMR7178, 25 Rue Becquerel, F-67087 Strasbourg, France. (author)
  • Lindskog, CeciliaUppsala universitet,Institutionen för immunologi, genetik och patologi,Science for Life Laboratory, SciLifeLab(Swepub:uu)cecli129 (author)
  • Freour, ThomasCHU Nantes, Serv Med Reprod, 38 Blvd Jean Monnet, F-44093 Nantes, France.;INSERM, UMR1064, F-44093 Nantes, France. (author)
  • Vandenbrouck, YvesCEA, DRF, BIG, Lab Biol Grande Echelle, 17 Rue Martyrs, F-38054 Grenoble, France.;INSERM, U1038, F-38054 Grenoble, France.;Grenoble Alpes Univ, F-38054 Grenoble, France. (author)
  • Lane, LydieSIB, CALIPHO Grp, CMU, Rue Michel Servet 1, CH-1211 Geneva 4, Switzerland.;Univ Geneva, Fac Med, Dept Human Prot Sci, 1 Rue Michel Servet, CH-1211 Geneva 4, Switzerland. (author)
  • Pineau, CharlesIrset, INSERM, U1085, Protim, Campus Beaulieu, F-35042 Rennes, France. (author)
  • 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. (creator_code:org_t)

Related titles

  • In:Journal of Proteome Research: American Chemical Society (ACS)16:12, s. 4340-43511535-38931535-3907

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