{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2024,7,18]],"date-time":"2024-07-18T07:45:03Z","timestamp":1721288703798},"reference-count":52,"publisher":"Wiley","issue":"4","license":[{"start":{"date-parts":[[2013,3,4]],"date-time":"2013-03-04T00:00:00Z","timestamp":1362355200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["J of Molecular Recognition"],"published-print":{"date-parts":[[2013,4]]},"abstract":"<jats:p>Protein\u2013protein interactions play central roles in physiological and pathological processes. The bases of the mechanisms of drug action are relevant to the discovery of new therapeutic targets. This work focuses on understanding the interactions in protein\u2013protein\u2013ligands complexes, using proteins calmodulin (CaM), human calcium\/calmodulin\u2010dependent 3\u2032,5\u2032\u2010cyclic nucleotide phosphodiesterase 1A active human (PDE1A), and myosin light chain kinase (MLCK) and ligands \u03b1II\u2013spectrin peptide (\u03b1II\u2013spec), and two inhibitors of CaM (chlorpromazine (CPZ) and malbrancheamide (MBC)). The interaction was monitored with a fluorescent biosensor of CaM (<jats:italic>h<\/jats:italic>CaM M124C\u2013<jats:italic>mBBr<\/jats:italic>). The results showed changes in the affinity of CPZ and MBC depending on the CaM\u2013protein complex under analysis. For the Ca<jats:sup>2+<\/jats:sup>\u2013CaM, Ca<jats:sup>2+<\/jats:sup>\u2013CaM\u2013PDE1A, and Ca<jats:sup>2+<\/jats:sup>\u2013CaM\u2013MLCK complexes, CPZ apparent dissociation constants (<jats:italic>K<\/jats:italic><jats:sub>ds<\/jats:sub>) were 1.11, 0.28, and 0.55\u2009\u03bcM, respectively; and for MBC <jats:italic>K<\/jats:italic><jats:sub>ds<\/jats:sub> were 1.43, 1.10, and 0.61\u2009\u03bcM, respectively. In competition experiments the addition of calmodulin binding peptide 1 (\u03b1II\u2013spec) to Ca<jats:sup>2+<\/jats:sup>\u2013<jats:italic>h<\/jats:italic>CaM M124C\u2013<jats:italic>mBBr<\/jats:italic> quenched the fluorescence (<jats:italic>K<\/jats:italic><jats:sub>d<\/jats:sub>\u2009=\u20092.55\u2009\u00b1\u20091.75\u2009pM) and the later addition of MBC (up to 16\u2009\u03bcM) did not affect the fluorescent signal. Instead, the additions of \u03b1II\u2013spec to a preformed Ca<jats:sup>2+<\/jats:sup>\u2013<jats:italic>h<\/jats:italic>CaM M124C\u2013<jats:italic>mBBr<\/jats:italic>\u2013MBC complex modified the fluorescent signal. However, MBC was able to displace the PDE1A and MLCK from its complex with Ca<jats:sup>2+<\/jats:sup>\u2013CaM. In addition, docking studies were performed for all complexes with both ligands showing an excellent correlation with experimental data. These experiments may help to explain why <jats:italic>in vivo<\/jats:italic> many CaM drugs target prefer only a subset of the Ca<jats:sup>2+<\/jats:sup>\u2013CaM regulated proteins and adds to the understanding of molecular interactions between protein complexes and small ligands. Copyright \u00a9 2013 John Wiley &amp; Sons, Ltd.<\/jats:p>","DOI":"10.1002\/jmr.2261","type":"journal-article","created":{"date-parts":[[2013,3,4]],"date-time":"2013-03-04T18:32:27Z","timestamp":1362421947000},"page":"165-174","source":"Crossref","is-referenced-by-count":5,"title":["Importance of the interaction protein\u2013protein of the CaM\u2013PDE1A and CaM\u2013MLCK complexes in the development of new anti\u2010CaM drugs"],"prefix":"10.1002","volume":"26","author":[{"given":"Martin","family":"Gonz\u00e1lez\u2010Andrade","sequence":"first","affiliation":[{"name":"Instituto Nacional de Medicina Gen\u00f3mica, Secretar\u00eda de Salud  M\u00e9xico DF 14610 M\u00e9xico"},{"name":"Facultad de Qu\u00edmica Universidad Nacional Aut\u00f3noma de M\u00e9xico  M\u00e9xico DF 04510 M\u00e9xico"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rachel","family":"Mata","sequence":"additional","affiliation":[{"name":"Facultad de Qu\u00edmica Universidad Nacional Aut\u00f3noma de M\u00e9xico  M\u00e9xico DF 04510 M\u00e9xico"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Abraham","family":"Madariaga\u2010Maz\u00f3n","sequence":"additional","affiliation":[{"name":"Facultad de Qu\u00edmica Universidad Nacional Aut\u00f3noma de M\u00e9xico  M\u00e9xico DF 04510 M\u00e9xico"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rogelio","family":"Rodr\u00edguez\u2010Sotres","sequence":"additional","affiliation":[{"name":"Facultad de Qu\u00edmica Universidad Nacional Aut\u00f3noma de M\u00e9xico  M\u00e9xico DF 04510 M\u00e9xico"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Luis","family":"del Pozo\u2010Yauner","sequence":"additional","affiliation":[{"name":"Instituto Nacional de Medicina Gen\u00f3mica, Secretar\u00eda de Salud  M\u00e9xico DF 14610 M\u00e9xico"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alejandro","family":"Sosa\u2010Peinado","sequence":"additional","affiliation":[{"name":"Facultad de Medicina Universidad Nacional Aut\u00f3noma de M\u00e9xico  M\u00e9xico DF 04510 M\u00e9xico"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2013,3,4]]},"reference":[{"key":"e_1_2_6_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0070-2137(01)80011-X"},{"key":"e_1_2_6_3_1","doi-asserted-by":"publisher","DOI":"10.1152\/physrev.1995.75.4.725"},{"key":"e_1_2_6_4_1","first-page":"200","article-title":"Characterization of the binding of calmodulin to non\u2010erythroid spectrin","volume":"66","author":"Bjork J","year":"1995","journal-title":"Eur. 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