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dc.creatorGrozdanović, Milica
dc.creatorDrakulić, Branko
dc.creatorGavrović-Jankulović, Marija
dc.date.accessioned2019-04-26T21:41:02Z
dc.date.available2019-04-26T21:41:02Z
dc.date.issued2013
dc.identifier.issn0304-4165
dc.identifier.urihttp://cer.ihtm.bg.ac.rs/handle/123456789/2714
dc.description.abstractBackground: Actinidin, a protease from kiwifruit, belongs to the C1 family of cysteine proteases. Cysteine proteases were found to be involved in many disease states and are valid therapeutic targets. Actinidin has a wide pH activity range and wide substrate specificity, which makes it a good model system for studying enzyme-substrate interactions. Methods: The influence of inhibitor (E-64) binding on the conformation of actinidin was examined by 2D PAGE, circular dichroism (CD) spectroscopy, hydrophobic ligand binding assay, and molecular dynamics simulations. Results: Significant differences were observed in electrophoretic mobility of proteolytically active and E-64-inhibited actinidin. CD spectrometry and hydrophobic ligand binding assay revealed a difference in conformation between active and inhibited actinidin. Molecular dynamics simulations showed that a loop defined by amino-acid residues 88-104 had greater conformational mobility in the inhibited enzyme than in the active one. During MD simulations, the covalently bound inhibitor was found to change its conformation from extended to folded, with the guanidino moiety approaching the carboxylate. Conclusions: Conformational mobility of actinidin changes upon binding of the inhibitor, leading to a sequence of events that enables water and ions to protrude into a newly formed cavity of the inhibited enzyme. Drastic conformational mobility of E-64, a common inhibitor of cysteine proteases found in many crystal structures stored in PDB, was also observed. General significance: The analysis of structural changes which occur upon binding of an inhibitor to a cysteine protease provides a valuable starting point for the future design of therapeutic agents. (c) 2013 Elsevier B.V. All rights reserved.en
dc.publisherElsevier Science Bv, Amsterdam
dc.relationinfo:eu-repo/grantAgreement/MESTD/Basic Research (BR or ON)/172049/RS//
dc.relationinfo:eu-repo/grantAgreement/MESTD/Basic Research (BR or ON)/172035/RS//
dc.relationinfo:eu-repo/grantAgreement/EC/FP7/256716/EU//
dc.relationinfo:eu-repo/grantAgreement/EC/FP7/261499/EU//
dc.rightsrestrictedAccess
dc.sourceBiochimica et Biophysica Acta: General Subjects
dc.subjectActinidinen
dc.subjectE-64en
dc.subjectMolecular dynamicsen
dc.subjectCysteine proteaseen
dc.titleConformational mobility of active and E-64-inhibited actinidinen
dc.typearticle
dc.rights.licenseARR
dcterms.abstractГавровић-Јанкуловић, Марија; Гроздановиц, Милица М.; Дракулић, Бранко;
dc.rights.holderElsevier
dc.citation.volume1830
dc.citation.issue10
dc.citation.spage4790
dc.citation.epage4799
dc.citation.other1830(10): 4790-4799
dc.citation.rankM21
dc.identifier.pmid23803410
dc.identifier.doi10.1016/j.bbagen.2013.06.015
dc.identifier.scopus2-s2.0-84880183998
dc.identifier.wos000323854900041
dc.type.versionpublishedVersionen


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