MspA Porin-Gold Nanoparticle Assemblies: Enhanced Binding through a Controlled Cysteine Mutation.
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.
Dani
Raj Kumar
RK
Department of Chemistry, Kansas State University, Manhattan, KS 66506-3701, USA.
Kang
Myungshim
M
Kalita
Mausam
M
Smith
Paul E
PE
Bossmann
Stefan H
SH
Chikan
Viktor
V
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
D018272:Porins; C121838:mspA protein, Mycobacterium smegmatis; D006046:Gold; D003545:Cysteine