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dc.contributor.authorYoung, Kenneth L.
dc.contributor.authorKassouf, Claudia
dc.contributor.authorDolinska, Monika B.
dc.contributor.authorAnderson, David Eric
dc.contributor.authorSergeev, Yuri V.
dc.date.accessioned2020-03-02T13:25:54Z
dc.date.available2020-03-02T13:25:54Z
dc.date.issued2020-01-30
dc.identifier.citationYoung, K.L., II; Kassouf, C.; Dolinska, M.B.; Anderson, D.E.; Sergeev, Y.V. Human Tyrosinase: Temperature-Dependent Kinetics of Oxidase Activity. Int. J. Mol. Sci. 2020, 21, 895.
dc.identifier.urihttp://hdl.handle.net/10919/97090
dc.description.abstractHuman tyrosinase (Tyr) is involved in pigment biosynthesis, where mutations in its corresponding gene TYR have been linked to oculocutaneous albinism 1, an autosomal recessive disorder. Although the enzymatic capabilities of Tyr have been well-characterized, the thermodynamic driving forces underlying melanogenesis remain unknown. Here, we analyze protein binding using the diphenol oxidase behavior of Tyr and van ’t Hoff temperature-dependent analysis. Recombinant Tyr was expressed and purified using a combination of affinity and size-exclusion chromatography. Michaelis-Menten constants were measured spectrophotometrically from diphenol oxidase reactions of Tyr, using L-3,4-dihydroxyphenylalanine (L-DOPA) as a substrate, at temperatures: 25, 31, 37, and 43 °C. Under the same conditions, the Tyr structure and the L-DOPA binding activity were simulated using 3 ns molecular dynamics and docking. The thermal Michaelis-Menten kinetics data were subjected to the van ‘t Hoff analysis and fitted with the computational model. The temperature-dependent analysis suggests that the association of L-DOPA with Tyr is a spontaneous enthalpy-driven reaction, which becomes unfavorable at the final step of dopachrome formation.en
dc.format.mimetypeapplication/pdf
dc.language.isoenen_US
dc.publisherMDPI
dc.rightsCreative Commons Attribution 4.0 International*
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/*
dc.titleHuman Tyrosinase: Temperature-Dependent Kinetics of Oxidase Activityen
dc.typeArticle - Refereeden_US
dc.date.updated2020-03-02T12:39:24Z
dc.title.serialInternational Journal of Molecular Science
dc.identifier.doihttps://doi.org/10.3390/ijms21030895
dc.type.dcmitypeText


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License: Creative Commons Attribution 4.0 International