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Native ESI Mass Spectrometry Can Help to Avoid Wrong Interpretations from Isothermal Titration Calorimetry in Difficult Situations

dc.contributor.authorWOLFF, Philippe
dc.contributor.authorDA VEIGA, Cyrielle
dc.contributor.authorENNIFAR, Eric
dc.contributor.authorBEC, Guillaume
hal.structure.identifierChimie et Biologie des Membranes et des Nanoobjets [CBMN]
dc.contributor.authorGUICHARD, Gilles
IDREF: 084339268
dc.contributor.authorBURNOUF, Dominique
dc.contributor.authorDUMAS, Philippe
dc.date.accessioned2020-07-08T08:44:05Z
dc.date.available2020-07-08T08:44:05Z
dc.date.issued2017-02
dc.identifier.issn1044-0305
dc.identifier.urihttps://oskar-bordeaux.fr/handle/20.500.12278/10185
dc.description.abstractEnWe studied by native ESI-MS the binding of various DNA-polymerase-derived peptides onto DNA-polymerase processivity rings from Escherichia coli, Pseudomonas aeruginosa, and Mycobacterium tuberculosis. These homodimeric rings present two equivalent specific binding sites, which leads to successive formation during a titration experiment of singly- and doubly occupied rings. By using the ESI-MS free-ring spectrum as a ruler, we derived by robust linear regression the fractions of the different ring species at each step of a titration experiment. These results led to accurate K-d values (from 0.03 to 0.5 mu M) along with the probability of peptide loss due to gas phase dissociation (GPD). We show that this good quality is due to the increased information content of a titration experiment with a homodimer. Isothermal titration calorimetry (ITC) led with the same binding model to K-d(ITC) values systematically higher than their ESI-MS counterparts and, often, to poor fit of the ITC curves. A processing with two competing modes of binding on the same site requiring determination of two (K-d, Delta H) pairs greatly improved the fits and yielded a second K-d(ITC) close to K-d(ESI-MS). The striking features are: (1) ITC detected a minor binding mode (similar to 20%) of 'low-affinity' that did not appear with ESI-MS; (2) the simplest processing of ITC data with only one (K-d, Delta H) pair led wrongly to the Kd of the low-affinity binding mode but to the Delta H of the high-affinity binding mode. Analogous misleading results might well exist in published data based on ITC experiments.
dc.title.enNative ESI Mass Spectrometry Can Help to Avoid Wrong Interpretations from Isothermal Titration Calorimetry in Difficult Situations
dc.typeArticle de revue
dc.identifier.doi10.1007/s13361-016-1534-6
dc.subject.halChimie/Matériaux
bordeaux.journalJournal of The American Society for Mass Spectrometry
bordeaux.page347-357
bordeaux.volume28
bordeaux.hal.laboratoriesInstitut de Chimie & de Biologie des Membranes & des Nano-objets (CBMN) - UMR 5248*
bordeaux.hal.laboratoriesInstitut de Chimie & de Biologie des Membranes & des Nano-objets (CBMN, UMR 5248)
bordeaux.issue2
bordeaux.institutionUniversité de Bordeaux
bordeaux.institutionBordeaux INP
bordeaux.COinSctx_ver=Z39.88-2004&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.jtitle=Journal%20of%20The%20American%20Society%20for%20Mass%20Spectrometry&rft.date=2017-02&rft.volume=28&rft.issue=2&rft.spage=347-357&rft.epage=347-357&rft.eissn=1044-0305&rft.issn=1044-0305&rft.au=WOLFF,%20Philippe&DA%20VEIGA,%20Cyrielle&ENNIFAR,%20Eric&BEC,%20Guillaume&GUICHARD,%20Gilles&rft.genre=article


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