dc.contributor.author | Naik, Subhashchandra | |
dc.contributor.author | Brock, Susan R. | |
dc.contributor.author | Akkaladevi, Narahari | |
dc.contributor.author | Tally, Jon | |
dc.contributor.author | Mcginn-Staub, Wesley | |
dc.contributor.author | Zhang, Na | |
dc.contributor.author | Gao, Philip | |
dc.contributor.author | Gogol, E. P. | |
dc.contributor.author | Pentelute, B. L. | |
dc.contributor.author | Collier, R. John | |
dc.contributor.author | Fisher, Mark T. | |
dc.date.accessioned | 2017-04-14T19:38:11Z | |
dc.date.available | 2017-04-14T19:38:11Z | |
dc.date.issued | 2013-09-17 | |
dc.identifier.citation | Naik, S., Brock, S., Akkaladevi, N., Tally, J., Mcginn-Straub, W., Zhang, N., … Fisher, M. T. (2013). Monitoring the kinetics of the pH driven transition of the anthrax toxin prepore to the pore by biolayer interferometry and surface plasmon resonance. Biochemistry, 52(37), 6335–6347. http://doi.org/10.1021/bi400705n | en_US |
dc.identifier.uri | http://hdl.handle.net/1808/23702 | |
dc.description.abstract | Domain 2 of the anthrax protective antigen (PA) prepore heptamer unfolds and refolds during endosome acidification to generate an extended 100 Å beta barrel pore that inserts into the endosomal membrane. The PA pore facilitates the pH dependent unfolding and translocation of bound toxin enzymic components, lethal factor (LF) and/or edema factor (EF), from the endosome into the cytoplasm. We constructed immobilized complexes of the prepore with the PA-binding domain of LF (LFN) to monitor the real-time prepore to pore kinetic transition using surface plasmon resonance (SPR) and bio-layer interferometry (BLI). The kinetics of this transition increased as the solution pH was decreased from pH 7.5 to pH 5.0, mirroring acidification of the endosome. Once transitioned, the LFN-PA pore complex was removed from the BLI biosensor tip and deposited onto EM grids, where the PA pore formation was confirmed by negative stain electron microscopy. When the soluble receptor domain (ANTRX2/CMG2) binds the immobilized PA prepore, the transition to the pore state was observed only after the pH was lowered to early or late endosomal pH conditions (5.5 to 5.0 respectively). Once the pore formed, the soluble receptor readily dissociated from the PA pore. Separate binding experiments with immobilized PA pores and soluble receptor indicate that the receptor has a weakened propensity to bind to the transitioned pore. This immobilized anthrax toxin platform can be used to identify or validate potential antimicrobial lead compounds capable of regulating and/or inhibiting anthrax toxin complex formation or pore transitions. | en_US |
dc.publisher | ACS | en_US |
dc.rights | Copyright © 2013 American Chemical Society | en_US |
dc.title | Monitoring the kinetics of the pH driven transition of the anthrax toxin prepore to the pore by biolayer interferometry and surface plasmon resonance | en_US |
dc.type | Article | en_US |
kusw.kuauthor | Zhang, Na | |
kusw.kuauthor | Gao, Philip | |
kusw.kudepartment | Protein Production Facility | en_US |
kusw.kudepartment | Higuchi Biosciences Center | en_US |
kusw.oanotes | Per SherpaRomeo on 04/14/2017: Author's Pre-print: grey tick subject to Restrictions below, author can archive pre-print (ie pre-refereeing) Restrictions: Must obtain written permission from Editor Must not violate ACS ethical Guidelines Author's Post-print: grey tick subject to Restrictions below, author can archive post-print (ie final draft post-refereeing) Restrictions: If mandated by funding agency or employer/ institution If mandated to deposit before 12 months, must obtain waiver from Institution/Funding agency or use AuthorChoice 12 months embargo Publisher's Version/PDF: cross author cannot archive publisher's version/PDF General Conditions: On author's personal website, pre-print servers, institutional website, institutional repositories or subject repositories Non-Commercial Must be accompanied by set statement (see policy) Must link to publisher version Publisher's version/PDF cannot be used | en_US |
dc.identifier.doi | 10.1021/bi400705n | en_US |
kusw.oaversion | Scholarly/refereed, author accepted manuscript | en_US |
kusw.oapolicy | This item meets KU Open Access policy criteria. | en_US |
dc.rights.accessrights | openAccess | |