{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,16]],"date-time":"2026-10-16T22:33:24Z","timestamp":1792190004763,"version":"4.4.0"},"reference-count":44,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2012,5,2]],"date-time":"2012-05-02T00:00:00Z","timestamp":1335916800000},"content-version":"vor","delay-in-days":1,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["J. Mass. Spectrom."],"published-print":{"date-parts":[[2012,5]]},"abstract":"<jats:p>DM43 is a circulating dimeric antitoxin isolated from <jats:italic>Didelphis aurita<\/jats:italic>, a South American marsupial naturally immune to snake envenomation. This endogenous inhibitor binds non\u2010covalently to jararhagin, the main hemorrhagic metalloproteinase from <jats:italic>Bothrops jararaca<\/jats:italic> snake venom, and efficiently neutralizes its toxicity. The aim of this study was to apply mass spectrometry (MS) and surface plasmon resonance (SPR) to improve the molecular characterization of this heterocomplex. The stoichiometry of the interaction was confirmed by nanoelectrospray ionization\u2010quadrupole\u2010time\u2010of\u2010flight MS; from native solution conditions, the complex showed a molecular mass of\u2009~94\u2009kDa, indicating that one molecule of jararhagin (50\u2009kDa) interacts with one monomer of DM43 (43\u2009kDa). Although readily observed in solution, the dimeric structure of the inhibitor was barely preserved in the gas phase. This result suggests that, in contrast to the toxin\u2013antitoxin complex, hydrophobic interactions are the primary driving force for the inhibitor dimerization. For the real\u2010time interaction analysis, the toxin was captured on a sensor chip derivatized with the anti\u2010jararhagin monoclonal antibody MAJar 2. The sensorgrams obtained after successive injections of DM43 in a concentration series were globally fitted to a simple bimolecular interaction, yielding the following kinetic rates for the DM43\/jararhagin interaction: <jats:italic>k<jats:sub>a<\/jats:sub><\/jats:italic>\u2009=\u20093.54 \u00b1 0.03 \u00d7 10<jats:sup>4<\/jats:sup>\u2009M<jats:sup>\u22121<\/jats:sup>\u2009s<jats:sup>\u22121<\/jats:sup> and <jats:italic>k<jats:sub>d<\/jats:sub><\/jats:italic>\u2009=\u20091.16 \u00b1 0.07 \u00d7 10<jats:sup>\u22125<\/jats:sup>\u2009s<jats:sup>\u22121<\/jats:sup>, resulting in an equilibrium dissociation constant (<jats:italic>K<jats:sub>D<\/jats:sub><\/jats:italic>) of 0.33\u2009\u00b1\u20090.06 nM. Taken together, MS and SPR results show that DM43 binds to its target toxin with high affinity and constitute the first accurate quantitative study on the extent of the interaction between a natural inhibitor and a metalloproteinase toxin, with unequivocal implications for the use of this kind of molecule as template for the rational development of novel antivenom therapies. Copyright \u00a9 2012 John Wiley &amp; Sons, Ltd.<\/jats:p>","DOI":"10.1002\/jms.2990","type":"journal-article","created":{"date-parts":[[2012,5,2]],"date-time":"2012-05-02T06:48:16Z","timestamp":1335941296000},"page":"567-573","source":"Crossref","is-referenced-by-count":12,"title":["The interaction of the antitoxin DM43 with a snake venom metalloproteinase analyzed by mass spectrometry and surface plasmon resonance"],"prefix":"10.1002","volume":"47","author":[{"given":"Guilherme D.","family":"Brand","sequence":"first","affiliation":[{"name":"Laborat\u00f3rio de Espectrometria de Massa Embrapa\u2010Recursos Gen\u00e9ticos e Biotecnologia, Esta\u00e7\u00e3o Parque Biol\u00f3gico  Final W5, Asa Norte 70770\u2010900 Bras\u00edlia DF Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rune","family":"Salbo","sequence":"additional","affiliation":[{"name":"Diabetes Protein Engineering Novo Nordisk A\/S  Novo Nordisk Park DK\u20102760 M\u00e5l\u00f8v Denmark"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thomas J. 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Vital Brazil 1500 Butant\u00e3 05503\u2010900 S\u00e3o Paulo, SP Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ana","family":"M. Moura\u2010da\u2010Silva","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Imunopatologia Instituto Butantan  Av. Vital Brazil 1500 Butant\u00e3 05503\u2010900 S\u00e3o Paulo, SP Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peter","family":"Roepstorff","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology University of Southern Denmark  Campusvej 55 DK\u20105230 Odense M Denmark"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gilberto B.","family":"Domont","sequence":"additional","affiliation":[{"name":"Unidade Prote\u00f4mica, Laborat\u00f3rio de Qu\u00edmica de Prote\u00ednas, Instituto de Qu\u00edmica, Centro de Tecnologia Universidade Federal do Rio de Janeiro  21941\u2010909 Rio de Janeiro RJ Brazil"},{"name":"Rio de Janeiro Proteomic Network\/FAPERJ"},{"name":"Instituto Nacional de Ci\u00eancia e Tecnologia em Toxinas\/CNPq"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jonas","family":"Perales","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Toxinologia, Pavilh\u00e3o Oz\u00f3rio de Almeida Instituto Oswaldo Cruz  Fiocruz, Av. 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