3 years ago

Using SiO2-Coated Gold Nanorods as Temperature Jump Photothermal Convertors Coupled with a Confocal Fluorescent Thermometer to Study Protein Unfolding Kinetics: A Case of Bovine Serum Albumin

Using SiO2-Coated Gold Nanorods as Temperature Jump Photothermal Convertors Coupled with a Confocal Fluorescent Thermometer to Study Protein Unfolding Kinetics: A Case of Bovine Serum Albumin
Chia-Te Lin, Ke-Chia Tseng, Li-Kang Chu, Kuan-Jen Chen
Photoexcitation of SiO2-coated gold nanorods (AuNR@SiO2) with a 1064 nm pulse of ca. 100 μs duration leads to a temperature jump of 5 °C, as determined by the evolution of the fluorescence intensity of tryptophan. A confocal fluorescent thermometer was developed to detect the temperature evolution in a small excitation volume of ca. 10–3 mm3 and provide a spatial resolution of 100 μm. In addition, a continuous-wave laser at 1550 nm was employed to increase the initial temperature to 44 °C by heating H2O, providing alternative initial temperatures for temperature jump experiments to reveal the unfolding kinetics of bovine serum albumin (BSA). The evolution of fluorescence of BSA upon temperature jump when the stationary temperature was raised to 44 °C differed from that at room temperature, suggesting a dynamical unfolding kinetics, and a rate coefficient of 75 ± 15 s–1 was derived. In this work, we successfully demonstrated the applicability of AuNR@SiO2 as the photothermal material for the temperature jump and employment of a confocal fluorescent thermometer to precisely monitor the minuscule heating volume. It would be advantageous to utilize this apparatus as an alternative tool for studying the kinetics of protein unfolding.

Publisher URL: http://dx.doi.org/10.1021/acs.jpcc.7b05033

DOI: 10.1021/acs.jpcc.7b05033

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