Catalytic importance of the substrate binding order for the FMNH2-dependent alkanesulfonate monooxygenase enzyme

Biochemistry
Xuanzhi ZhanHolly R Ellis

Abstract

The two-component alkanesulfonate monooxygenase system from Escherichia coli includes an FMN reductase (SsuE) and an FMNH2-dependent alkanesulfonate monooxygenase (SsuD) involved in the acquisition of sulfur from alkanesulfonates during sulfur starvation. The SsuD enzyme directly catalyzes the oxidation of alkanesulfonate to aldehyde and sulfite in the presence of O2 and FMNH2. The goal of these studies was to investigate the kinetic mechanism of SsuD through rapid reaction kinetics and substrate binding studies. The SsuD enzyme shows a clear preference for FMNH2 (Kd, 0.32 +/- 0.15 microM) compared to FMN (Kd, 10.2 +/- 0.4 microM) with a 1:1 binding stoichiometry for each form of the flavin. The kinetic trace of premixed SsuD and FMNH2 mixed with oxygenated buffer was best fit to a double exponential with no observed formation of the C4a-(hydro)peroxyflavin. However, when FMNH2 was mixed with SsuD and oxygenated buffer an initial fast phase (kobs, 12.9 s-1) was observed, suggesting that the mixing order is critical for the accumulation of the C4a-(hydro)peroxyflavin. Results from fluorimetric titrations with octanesulfonate imply that reduced flavin must bind first to promote octanesulfonate binding. When octanesulfonate was in...Continue Reading

References

Jan 5, 1996·The Journal of Biological Chemistry·W A FranciscoF M Raushel
Sep 12, 1997·The Journal of Biological Chemistry·E EichhornT Leisinger
Sep 10, 1999·The Journal of Biological Chemistry·E EichhornT Leisinger
Aug 2, 2001·Archives of Microbiology·J R van der PloegT Leisinger
Nov 26, 2002·Journal of Molecular Biology·Eric EichhornTimothy J Richmond
Aug 7, 2004·The Journal of Biological Chemistry·Julien ValtonVincent Nivière
Sep 9, 2004·The Biochemical Journal·Saman HosseinkhaniEdward A Meighen

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Citations

Dec 5, 2015·Biochemistry·Paritosh V DayalHolly R Ellis
Mar 3, 2010·Archives of Biochemistry and Biophysics·Holly R Ellis
Sep 2, 2018·Protein Science : a Publication of the Protein Society·Jeffrey S McFarlaneHolly R Ellis
Jun 7, 2020·Applied and Environmental Microbiology·Chulwoo ParkWoojun Park
Feb 6, 2017·The Journal of Biological Chemistry·Panu PimviriyakulPimchai Chaiyen
Dec 15, 2018·Polish Journal of Microbiology·DA Cheng HaoPei Gen Xiao
Nov 15, 2020·The Journal of Biological Chemistry·Aisaraphon PhinthaPimchai Chaiyen
Jan 20, 2021·The Journal of Biological Chemistry·Aisaraphon PhinthaPimchai Chaiyen

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