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dc.contributor.authorMachovina, Melodie M.
dc.contributor.authorEllis, Emerald S.
dc.contributor.authorCarney, Thomas J.
dc.contributor.authorBrushett, Fikile R.
dc.contributor.authorDuBois, Jennifer L.
dc.date.accessioned2019-03-05T15:33:14Z
dc.date.available2019-03-05T15:33:14Z
dc.date.issued2019-01
dc.identifier.citationMachovina, Melodie M., Emerald S Ellis, Thomas J Carney, Fikile R. Brushett, and Jennifer L. DuBois. "How a cofactor-free protein environment lowers the barrier to O2 reactivity." Journal of Biological Chemistry (January 2019). DOI:10.1074/jbc.RA118.006144.en_US
dc.identifier.issn1083-351X
dc.identifier.urihttps://scholarworks.montana.edu/xmlui/handle/1/15311
dc.description.abstractMolecular oxygen (O2)-utilizing enzymes are among the most important in biology. The abundance of O2, its thermodynamic power, and the benign nature of its end products have raised interest in oxidases and oxygenases for biotechnological applications. While most O2-dependent enzymes have an absolute requirement for an O2-activating cofactor, several classes of oxidases and oxygenases accelerate direct reactions between substrate and O2 using only the protein environment. Nogalamycin monooxygenase (NMO) from Streptomyces nogalater is a cofactor-independent enzyme that catalyzes rate-limiting electron transfer between its substrate and O2. Here, using enzyme-kinetic, cyclic voltammetry, and mutagenesis methods, we demonstrate that NMO initially activates the substrate, lowering its pKa by 1.0 unit (ΔG*= 1.4 kcal mol-1). We found that the one-electron reduction potential, measured for the deprotonated substrate both inside and outside the protein environment, increases by 85 mV inside NMO, corresponding to a ΔΔG⁰′ of 2.0 kcal mol-1 (0.087 eV) and that the activation barrier, ΔG‡, is lowered by 4.8 kcal mol-1 (0.21 eV). Applying the Marcus model, we observed that this suggests a sizable decrease of 28 kcal mol-1 (1.4 eV) in the reorganization energy (l), which constitutes the major portion of the protein environment’s effect in lowering the reaction barrier. A similar role for the protein has been proposed in several cofactor-dependent systems and may reflect a broader trend in O2-utilizing proteins. In summary, NMO’s protein environment facilitates direct electron transfer, and NMO accelerates rate-limiting electron transfer by strongly lowering the reorganization energy.en_US
dc.description.sponsorshipNSF Grant MCB1715176en_US
dc.language.isoenen_US
dc.rightsThis Item is protected by copyright and/or related rights. You are free to use this Item in any way that is permitted by the copyright and related rights legislation that applies to your use. For other uses you need to obtain permission from the rights-holder(s).en_US
dc.rights.urihttp://rightsstatements.org/vocab/InC/1.0/en_US
dc.titleHow a cofactor-free protein environment lowers the barrier to O2 reactivityen_US
dc.typeArticleen_US
mus.citation.journaltitleJournal of Biological Chemistryen_US
mus.identifier.categoryChemical & Material Sciencesen_US
mus.identifier.doi10.1074/jbc.RA118.006144en_US
mus.relation.collegeCollege of Letters & Scienceen_US
mus.relation.departmentChemistry & Biochemistry.en_US
mus.relation.universityMontana State University - Bozemanen_US
mus.data.thumbpage14en_US


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