Human Tyrosinase: Temperature-Dependent Kinetics of Oxidase Activity
| dc.contributor.author | Young, Kenneth L. | en |
| dc.contributor.author | Kassouf, Claudia | en |
| dc.contributor.author | Dolinska, Monika B. | en |
| dc.contributor.author | Anderson, David Eric | en |
| dc.contributor.author | Sergeev, Yuri V. | en |
| dc.date.accessioned | 2020-03-02T13:25:54Z | en |
| dc.date.available | 2020-03-02T13:25:54Z | en |
| dc.date.issued | 2020-01-30 | en |
| dc.date.updated | 2020-03-02T12:39:24Z | en |
| dc.description.abstract | Human tyrosinase (Tyr) is involved in pigment biosynthesis, where mutations in its corresponding gene <i>TYR</i> 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.description.version | Published version | en |
| dc.format.mimetype | application/pdf | en |
| dc.identifier.citation | Young, 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. | en |
| dc.identifier.doi | https://doi.org/10.3390/ijms21030895 | en |
| dc.identifier.uri | http://hdl.handle.net/10919/97090 | en |
| dc.language.iso | en | en |
| dc.publisher | MDPI | en |
| dc.rights | Creative Commons Attribution 4.0 International | en |
| dc.rights.uri | http://creativecommons.org/licenses/by/4.0/ | en |
| dc.subject | tyrosinase | en |
| dc.subject | protein purification | en |
| dc.subject | L-DOPA binding | en |
| dc.subject | enthalpy-driven association | en |
| dc.title | Human Tyrosinase: Temperature-Dependent Kinetics of Oxidase Activity | en |
| dc.title.serial | International Journal of Molecular Science | en |
| dc.type | Article - Refereed | en |
| dc.type.dcmitype | Text | en |