Cross-Species Analysis of Protein Dynamics Associated with Hydride and Proton Transfer in the Catalytic Cycle of the Light-Driven Enzyme Protochlorophyllide Oxidoreductase.

Biochemistry
Robin HoevenNigel S Scrutton

Abstract

Experimental interrogation of the relationship between protein dynamics and enzyme catalysis is challenging. Light-activated protochlorophyllide oxidoreductase (POR) is an excellent model for investigating this relationship because photoinitiation of the reaction cycle enables coordinated turnover in a "dark-assembled" ternary enzyme-substrate complex. The catalytic cycle involves sequential hydride and proton transfers (from NADPH and an active site tyrosine residue, respectively) to the substrate protochlorophyllide. Studies with a limited cross-species subset of POR enzymes (n = 4) have suggested that protein dynamics associated with hydride and proton transfer are distinct [Heyes, D. J., Levy, C., Sakuma, M., Robertson, D. L., and Scrutton, N. S. (2011) J. Biol. Chem. 286, 11849-11854]. Here, we use steady-state assays and single-turnover laser flash spectroscopy to analyze hydride and proton transfer dynamics in an extended series of POR enzymes taken from many species, including cyanobacteria, algae, embryophytes, and angiosperms. Hydride/proton transfer in all eukaryotic PORs is faster compared to prokaryotic PORs, suggesting active site architecture has been optimized in eukaryotic PORs following endosymbiosis. Visible ...Continue Reading

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Citations

Jun 11, 2016·Journal of Photochemistry and Photobiology. B, Biology·Binuraj R K MenonDerren J Heyes
Jan 31, 2018·Protein Science : a Publication of the Protein Society·David D BoehrKathleen F O'Rourke
Oct 28, 2019·Nature·Shaowei ZhangNigel S Scrutton
Apr 3, 2020·Proceedings of the National Academy of Sciences of the United States of America·Chen-Song DongLin Liu
Mar 10, 2021·Nature Plants·Derren J HeyesNigel S Scrutton
Jul 13, 2018·The Journal of Physical Chemistry. B·Samira GholamiMarco Garavelli

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