title: MspA Porin-Gold Nanoparticle Assemblies: Enhanced Binding through a Controlled Cysteine Mutation.
abstract: In this study, the interactions of two gold nanoparticles of different sizes (average diameters of 3.7 +/- 2.6 and 17 +/- 3 nm) with octameric mycobacterial porin A from Mycobacterium smegmatis (MspA) and a mutant of MspA featuring a cysteine mutation in position 126 (Q126C) are investigated. From the observation of enhanced photoluminescence quenching, it is inferred that the presence of eight cysteines in the MspA Q126C mutant significantly enhances the binding of selected small gold nanoparticles within the inner pore of MspA. The large gold nanoparticle/porin complex shows photoluminescence enhancement, which is expected since the larger nanoparticles cannot dock within the homopore of MspA due to size exclusion. In addition to the fluorescence experiments, observation of energy transfer from the small gold nanoparticles to the MspA shows the close proximity of the small gold nanoparticles with the porin. Interestingly, the energy transfer of the large nanoparticle/MspA complex is completely missing. From high-performance liquid chromatography data, the estimated binding constants for small Au@MspA, large Au@MspA, small Au@MspAcys, and large Au@MspAcys are 1.3 x 10 (9), 2.22 x 10 (10), > 10 (12) (irreversible), and 1.7 x 10 (10), respectively.
authors.affiliation: Department of Chemistry, Kansas State University, Manhattan, KS 66506-3701, USA.
authors.forename: Raj Kumar
authors.initials: RK
authors.lastname: Dani
authors.forename: Myungshim
authors.initials: M
authors.lastname: Kang
authors.forename: Mausam
authors.initials: M
authors.lastname: Kalita
authors.forename: Paul E
authors.initials: PE
authors.lastname: Smith
authors.forename: Stefan H
authors.initials: SH
authors.lastname: Bossmann
authors.forename: Viktor
authors.initials: V
authors.lastname: Chikan
mesh_terms: D002851:Chromatography, High Pressure Liquid; D003545:Cysteine / Q000235:genetics*; D006046:Gold / Q000737:chemistry*; D053768:Metal Nanoparticles; D046529:Microscopy, Electron, Transmission; D009154:Mutation; D009501:Neutron Activation Analysis; D018272:Porins / Q000737:chemistry*; D013050:Spectrometry, Fluorescence
chemical_list: D018272:Porins; C121838:mspA protein, Mycobacterium smegmatis; D006046:Gold; D003545:Cysteine