Quantitative Peptidomics with Five-plex Reductive Methylation labels

Página do item simplificado
dc.citation.issue5
dc.citation.volumev. 29
dc.contributor.authorTashima, Alexandre Keiji [UNIFESP]
dc.contributor.authorFricker, Lloyd D.
dc.coverageNew York
dc.date.accessioned2020-07-20T16:30:52Z
dc.date.available2020-07-20T16:30:52Z
dc.date.issued2018
dc.description.abstractQuantitative peptidomics and proteomics often use chemical tags to covalently modify peptides with reagents that differ in the number of stable isotopes, allowing for quantitation of the relative peptide levels in the original sample based on the peak height of each isotopic form. Different chemical reagents have been used as tags for quantitative peptidomics and proteomics, and all have strengths and weaknesses. One of the simplest approaches uses formaldehyde and sodium cyanoborohydride to methylate amines, converting primary and secondary amines into tertiary amines. Up to five different isotopic forms can be generated, depending on the isotopic forms of formaldehyde and cyanoborohydride reagents, allowing for five-plex quantitation. However, the mass difference between each of these forms is only 1 Da per methyl group incorporated into the peptide, and for many peptides there is substantial overlap from the natural abundance of C-13 and other isotopes. In this study, we calculated the contribution from the natural isotopes for 26 native peptides and derived equations to correct the peak intensities. These equations were applied to data from a study using human embryonic kidney HEK293T cells in which five replicates were treated with 100 nM vinblastine for 3 h and compared with five replicates of cells treated with control medium. The correction equations brought the replicates to the expected 1:1 ratios and revealed significant decreases in levels of 21 peptides upon vinblastine treatment. These equations enable accurate quantitation of small changes in peptide levels using the reductive methylation labeling approach.en
dc.description.affiliationUniv Fed Sao Paulo, Escola Paulista Med, Dept Biochem, BR-04023901 Sao Paulo, SP, Brazil
dc.description.affiliationAlbert Einstein Coll Med, Dept Mol Pharmacol, Bronx, NY 10461 USA
dc.description.affiliationAlbert Einstein Coll Med, Dept Neurosci, Bronx, NY 10461 USA
dc.description.sourceWeb of Science
dc.description.sponsorshipFinanciadora de Estudos e Projetos
dc.description.sponsorshipFundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP)
dc.description.sponsorshipIDFAPESP: 2012/19321-9
dc.description.sponsorshipIDFAPESP: 2016/03839-0
dc.format.extent866-878
dc.identifierhttp://dx.doi.org/10.1007/s13361-017-1852-3
dc.identifier.citationJournal Of The American Society For Mass Spectrometry. New York, v. 29, n. 5, p. 866-878, 2018.
dc.identifier.doi10.1007/s13361-017-1852-3
dc.identifier.issn1044-0305
dc.identifier.urihttps://repositorio.unifesp.br/handle/11600/55567
dc.identifier.wosWOS:000431802500008
dc.language.isoeng
dc.publisherSpringer
dc.relation.ispartofJournal Of The American Society For Mass Spectrometry
dc.rightsinfo:eu-repo/semantics/openAccess
dc.subjectIsotopic labelsen
dc.subjectFormaldehydeen
dc.subjectCyanoborohydrideen
dc.titleQuantitative Peptidomics with Five-plex Reductive Methylation labelsen
dc.typeinfo:eu-repo/semantics/article
Arquivos
Coleções
EPM - Artigos