Origin of the anomalous circular dichroism spectra of many apomyoglobin mutants

dc.creatorRibeiro, EA
dc.creatorRamos, CHI
dc.date2004
dc.date42156
dc.date2014-11-14T04:23:13Z
dc.date2015-11-26T17:13:16Z
dc.date2014-11-14T04:23:13Z
dc.date2015-11-26T17:13:16Z
dc.date.accessioned2018-03-29T00:01:38Z
dc.date.available2018-03-29T00:01:38Z
dc.identifierAnalytical Biochemistry. Academic Press Inc Elsevier Science, v. 329, n. 2, n. 300, n. 306, 2004.
dc.identifier0003-2697
dc.identifierWOS:000221824500018
dc.identifier10.1016/j.ab.2004.01.016
dc.identifierhttp://www.repositorio.unicamp.br/jspui/handle/REPOSIP/69771
dc.identifierhttp://www.repositorio.unicamp.br/handle/REPOSIP/69771
dc.identifierhttp://repositorio.unicamp.br/jspui/handle/REPOSIP/69771
dc.identifier.urihttp://repositorioslatinoamericanos.uchile.cl/handle/2250/1281530
dc.descriptionSeveral authors have reported that many sperm whale apomyoglobin mutants show anomalous circular dichroism spectra. These mutants have a low molar ellipticity compared to the wild-type protein but in several cases have the same stability of unfolding. A model in which native apomyoglobin is not folded in the same manner as that in other proteins and in which mutants show progressive reductions in their degree of folding has been suggested to explain this phenomenon. However, nuclear magnetic resonance of the native apomyoglobin conformation has shown that this state is folded and compact, raising the possibility that the anomalous circular dichroism spectra could have another explanation. We studied several mutants with anomalous circular dichroism spectra and found that these proteins were all contaminated with nucleic acid that contributed to the ultraviolet absorption and caused uncertainty in the determination of protein concentration. The resulting overestimation of the concentration of apomyoglobin explains the phenomenon of anomalous circular dichroism spectra. We describe a procedure to remove the contaminant nucleic acid which yields accurate protein concentration measurements and provides the normal circular dichroism spectra. Our findings support a well-structured native conformation for apomyoglobin and may also be of the interest to scientists working with the purification of recombinant proteins. (C) 2004 Elsevier Inc. All rights reserved.
dc.description329
dc.description2
dc.description300
dc.description306
dc.languageen
dc.publisherAcademic Press Inc Elsevier Science
dc.publisherSan Diego
dc.publisherEUA
dc.relationAnalytical Biochemistry
dc.relationAnal. Biochem.
dc.rightsfechado
dc.rightshttp://www.elsevier.com/about/open-access/open-access-policies/article-posting-policy
dc.sourceWeb of Science
dc.subjectapomyoglobin
dc.subjectellipticity
dc.subjectprotein folding
dc.subjectmolten globule
dc.subjectspectroscopy
dc.subjectprotein purification
dc.subject4 Folding Intermediate
dc.subjectMolten Globule Intermediate
dc.subjectSperm-whale Apomyoglobin
dc.subjectProtein
dc.subjectMyoglobin
dc.subjectStability
dc.subjectPh
dc.subjectSubstitutions
dc.subjectDenaturation
dc.subjectAggregation
dc.titleOrigin of the anomalous circular dichroism spectra of many apomyoglobin mutants
dc.typeArtículos de revistas


Este ítem pertenece a la siguiente institución