zhpsЖурнал прикладной спектроскопииZhurnal Prikladnoii Spektroskopii0514-7506B. I. Stepanov Institute of Physics of the National Academy of Scienceszhps-169Research ArticleСтатьиАКТИВНОСТЬ, СТАБИЛЬНОСТЬ И СТРУКТУРА ПРИРОДНОЙ И МОДИФИЦИРОВАННОЙ K-РЕАГЕНТОМ ВУДВОРДА ГРИБНОЙ ТИРОЗИНАЗЫACTIVITY, STABILITY, AND STRUCTURE OF NATIVE AND MODIFIED BY WOODWARD REAGENT K MUSHROOM TYROSINASEEmamiS. .EmamiS. .noemail@neicon.ruPiriH. .PiriH. .noemail@neicon.ruGheibiN. .GheibiN. .ngheibi@qums.ac.irНаучно-исследовательский филиал Исламского университета АзадIslamic Azad UniversityЦентр исследования молекул и клеток, Казвинский университет медицинских наукCellular and Molecular Research Center, Qazvin University of Medical Sciences201710032020846969976Copyright © Emami S..., Piri H..., Gheibi N..., 20202020Emami S..., Piri H..., Gheibi N...Emami S..., Piri H..., Gheibi N...Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://zhps.ejournal.by/jour/article/view/169

Исследованы активность, структура и стабильность природных и модифицированных K-реагентом Вудворда (WRK) карбоновых остатков грибной тирозиназы (MT). Относительная активность уменьшилась с 100% для природного фермента до 77.9, 53.8, 39.4 и 26.4 % для его модификаций при соотношении [WRK]/[MT] = 2.5, 5, 25 и 50 соответственно. По кривым термической денатурации для природных и модифицированных ферментов вычислено Tm = 61.2, 60.1, 58.3, 53.9 и 45.5 °C. Уменьшение ∆GH2O при химической денатурации модифицированного фермента указывает на его нестабильность. Структурное исследование методами кругового дихроизма и собственной флуоресценции выявило флуктуацию вторичных и третичных структур МТ.

Mushroom tyrosinase (MT) was considered a good model for studying the inhibition, activation, and mutation of tyrosinase as the key enzyme of melanogenesis. In the present study, the activity, structure, reduction, and stability of native and modified enzymes were investigated after the modification of MT carboxylic residues by the Woodward reagent K (WRK). The relative activity of the sole enzyme was reduced from 100 to 77.9, 53.8, 39.4, and 26.4% after its modification by 2.5, 5, 25, and 50 ratios of [WRK]/[MT], respectively. The Tm values were calculated from thermal denaturation curves at 61.2, 60.1, 58.3, 53.9, and 45.5°C for the sole and modified enzymes. The reduction of the ∆GH2O values for the modified enzyme in chemical denaturation indicated instability. A structural study by CD and intrinsic fluorescence technique revealed the fluctuation of the secondary and tertiary structures of MT.

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The authors declare that there are no conflicts of interest present.