Abstract
Utilising the size homogeneity of nanodisc, we investigated the effect of lipid composition on the function of bacteriorhodopsin. The protein was reconstituted into nanodisc composed of different molar ratios of zwitterionic and negatively charged lipid. The characterization of bacteriorhodopsin’s photocycle kinetics was probed using transient absorption spectroscopy, where the photocycle was initiated using a pulsed laser. The results suggested that the photocycle recovery rate of bacteriorhodopsin is significantly decreased with a decrease of negatively charged lipid content, and photocycle intermediates N and O were shown to decreased as well. While the charge of the lipid hydrophilic head appeared to significantly alter the photocycle kinetics of bacteriorhodopsin, the structure of the lipid hydrophobic tail can also influence the photocycle. On the other hand, transient photocurrent measurement results suggested that lipid may play a role in the translocation of proton during photocycle activity.
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Acknowledgements
The results of this chapter were in collaboration with Professor Li-Kang Chu’s students from National Tsing Hua University, Hsinchu, Taiwan. In particular, the preparation of monomeric bR in Triton X-100, the steady state absorption spectroscopy, and the single-wavelength transient absorption spectroscopy were performed either by Tsung-Yen Lee or Yin Hsin. The full-wavelength time resolved difference spectra were recorded by Tsung-Yen Lee, and the electrochemical measurement was carried out by Yin Hsin.
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Yeh, V. (2019). Effect of Lipid Composition of Nanodisc. In: Study of Bacteriorhodopsin in a Controlled Lipid Environment. Springer Theses. Springer, Singapore. https://doi.org/10.1007/978-981-13-1238-0_3
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DOI: https://doi.org/10.1007/978-981-13-1238-0_3
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