{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,13]],"date-time":"2026-03-13T14:07:55Z","timestamp":1773410875875,"version":"3.50.1"},"reference-count":21,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2020,1,30]],"date-time":"2020-01-30T00:00:00Z","timestamp":1580342400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000009","name":"Foundation for the National Institutes of Health","doi-asserted-by":"publisher","award":["ZIA EY000476\u201010"],"award-info":[{"award-number":["ZIA EY000476\u201010"]}],"id":[{"id":"10.13039\/100000009","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Human 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 \u2019t 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 \u00b0C. 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 \u2018t 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.<\/jats:p>","DOI":"10.3390\/ijms21030895","type":"journal-article","created":{"date-parts":[[2020,2,3]],"date-time":"2020-02-03T01:25:51Z","timestamp":1580693151000},"page":"895","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":16,"title":["Human Tyrosinase: Temperature-Dependent Kinetics of Oxidase Activity"],"prefix":"10.3390","volume":"21","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3950-1921","authenticated-orcid":false,"suffix":"II","given":"Kenneth L.","family":"Young","sequence":"first","affiliation":[{"name":"National Eye Institute, National Institutes of Health, 31 Center Drive MSC 2510, Bethesda, MD 20892, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Claudia","family":"Kassouf","sequence":"additional","affiliation":[{"name":"National Eye Institute, National Institutes of Health, 31 Center Drive MSC 2510, Bethesda, MD 20892, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Monika B.","family":"Dolinska","sequence":"additional","affiliation":[{"name":"National Eye Institute, National Institutes of Health, 31 Center Drive MSC 2510, Bethesda, MD 20892, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"David Eric","family":"Anderson","sequence":"additional","affiliation":[{"name":"National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, 6 Center Dr. MSC2775, Bethesda, MD 20892, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7204-6572","authenticated-orcid":false,"given":"Yuri V.","family":"Sergeev","sequence":"additional","affiliation":[{"name":"National Eye Institute, National Institutes of Health, 31 Center Drive MSC 2510, Bethesda, MD 20892, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2020,1,30]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"41","DOI":"10.1111\/pcmr.12546","article-title":"Oculocutaneous albinism type 1: Link between mutations, tyrosinase conformational stability, and enzymatic activity","volume":"30","author":"Dolinska","year":"2017","journal-title":"Pigment Cell Melanoma Res."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"307","DOI":"10.1034\/j.1600-0749.2003.00045.x","article-title":"Oculocutaneous albinism type 1: The last 100 years","volume":"16","author":"Oetting","year":"2003","journal-title":"Pigment Cell Melanoma Res."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"21682","DOI":"10.1074\/jbc.M603841200","article-title":"Productive folding of tyrosinase ectodomain is controlled by the transmembrane anchor","volume":"281","author":"Popescu","year":"2006","journal-title":"J. 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