Characterization of Temperature-Dependent Kinetics of Oculocutaneous Albinism-Causing Mutants of Tyrosinase
| dc.contributor.author | Wachamo, Samuel A. | en |
| dc.contributor.author | Patel, Milan H. | en |
| dc.contributor.author | Varghese, Paul K. | en |
| dc.contributor.author | Dolinska, Monika B. | en |
| dc.contributor.author | Sergeev, Yuri V. | en |
| dc.date.accessioned | 2021-07-23T17:29:30Z | en |
| dc.date.available | 2021-07-23T17:29:30Z | en |
| dc.date.issued | 2021-07-21 | en |
| dc.date.updated | 2021-07-23T13:27:57Z | en |
| dc.description.abstract | Human tyrosinase (Tyr) is a glycoenzyme that catalyzes the first and rate-limiting step in melanin production, and its gene (<i>TYR</i>) is mutated in many cases of oculocutaneous albinism type phenotype in patients with OCA1 have only began to be examined and remain to be delineated. Here, we analyze the temperature-dependent kinetics of wild-type Tyr (WT) and two OCA1B mutant variants (R422Q and P406L) using Michaelis–Menten and Van’t Hoff analyses. Recombinant truncated human Tyr proteins (residues 19–469) were produced in the whole insect <i>Trichoplusia Ni</i> larvae. Proteins were purified by a combination of affinity and size-exclusion chromatography. The temperature dependence of diphenol oxidase protein activities and kinetic parameters were measured by dopachrome absorption. Using the same experimental conditions, computational simulations were performed to assess the temperature-dependent association of L-DOPA and Tyr. Our results revealed, for the first time, that the association of L-DOPA with R422Q and P406L followed by dopachrome formation is a complex reaction supported by enthalpy and entropy forces. We show that the WT has a higher turnover number as compared with both R422Q and P406L. Elucidating the kinetics and thermodynamics of mutant variants of Tyr in OCA1B helps to understand the mechanisms by which they lower Tyr catalytic activity and to discover novel therapies for patients. | en |
| dc.description.version | Published version | en |
| dc.format.mimetype | application/pdf | en |
| dc.identifier.citation | Wachamo, S.A.; Patel, M.H.; Varghese, P.K.; Dolinska, M.B.; Sergeev, Y.V. Characterization of Temperature-Dependent Kinetics of Oculocutaneous Albinism-Causing Mutants of Tyrosinase. Int. J. Mol. Sci. 2021, 22, 7771. | en |
| dc.identifier.doi | https://doi.org/10.3390/ijms22157771 | en |
| dc.identifier.uri | http://hdl.handle.net/10919/104381 | 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 | human recombinant tyrosinases | en |
| dc.subject | OCA1B mutations | en |
| dc.subject | temperature-dependent kinetics | en |
| dc.subject | Van’t Hoff analysis | en |
| dc.subject | L-DOPA binding | en |
| dc.subject | decreased dopachrome in mutant variants | en |
| dc.title | Characterization of Temperature-Dependent Kinetics of Oculocutaneous Albinism-Causing Mutants of Tyrosinase | en |
| dc.title.serial | International Journal of Molecular Science | en |
| dc.type | Article - Refereed | en |
| dc.type.dcmitype | Text | en |