Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/50331
DC FieldValueLanguage
dc.contributor.authorDoong, S.R.en_US
dc.contributor.author王敏盈zh_TW
dc.contributor.authorChen, Y.H.en_US
dc.contributor.authorLai, S.Y.en_US
dc.contributor.authorLee, C.C.en_US
dc.contributor.authorLin, Y.C.en_US
dc.contributor.authorWang, M.Y.en_US
dc.date2007zh_TW
dc.date.accessioned2014-06-06T08:49:20Z-
dc.date.available2014-06-06T08:49:20Z-
dc.identifier.issn0003-2700zh_TW
dc.identifier.urihttp://hdl.handle.net/11455/50331-
dc.description.abstractVP2, the single outer protein of infectious bursal disease virus capsid, can self-assemble into T = 1 subviral particle (SVP), which can be efficiently purified by immobilized metal ion affinity chromatography (IMAC). In this study, a systemic investigation of the adsorption behavior of VP2 SVP on Ni-NTA resin was performed to identify that His253 and His249 on the surface of SVP are the key factors accounted for the strong and heterogeneous interaction. First, an untagged VP2-441 SVP was constructed, expressed, and purified by IMAC to demonstrate that SVP can interact with immobilized Ni2+ ions on NTA resin without an inserted His tag. Second, equilibrium adsorption studies were used to demonstrate that SVP has a higher affinity to the immobilized Ni2+ ions than a model protein, bovine serum albumin, although the maximum amount of SVP bound per volume resin is limited by the pore size of the resin as verified by confocal microscopic analysis. Third, based on structural analysis and computer modeling, His253 and His249 on the surface of SVP are responsible for a strong heterogeneous and multiple adsorption with the immobilized Ni2+ ions; and this was confirmed by a point-mutation experiment. This is the first example to elucidate the interaction between the immobilized metal ions and viral particles at molecular level. A detailed understanding of SVP-immobilized metal ion interactions can provide useful strategies for engineering icosahedral protein nanoparticles to achieve a simple and one-step purification by IMAC.en_US
dc.language.isoen_USzh_TW
dc.relationAnalytical Chemistryen_US
dc.relation.ispartofseriesAnalytical Chemistry, Volume 79, Issue 20, Page(s) 7654-7661.en_US
dc.relation.urihttp://dx.doi.org/10.1021/ac070745oen_US
dc.subjectvirus-like particlesen_US
dc.subjectaffinity-chromatographyen_US
dc.subjectexchange chromatographyen_US
dc.subjectprotein adsorptionen_US
dc.subjectcrystal-structureen_US
dc.subjectescherichia-colien_US
dc.subjectcytochromeen_US
dc.subjectb(5)en_US
dc.subjectcapsid proteinen_US
dc.subjectamino-acidsen_US
dc.subjectpurificationen_US
dc.titleStrong and heterogeneous adsorption of infectious bursal disease VP2 subviral particle with immobilized metal ions dependent on two surface histidine residuesen_US
dc.typeJournal Articlezh_TW
dc.identifier.doi10.1021/ac070745ozh_TW
item.cerifentitytypePublications-
item.grantfulltextnone-
item.languageiso639-1en_US-
item.fulltextno fulltext-
item.openairecristypehttp://purl.org/coar/resource_type/c_18cf-
item.openairetypeJournal Article-
Appears in Collections:生物科技學研究所
Show simple item record
 

Google ScholarTM

Check

Altmetric

Altmetric


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.