|
ИСТИНА |
Войти в систему Регистрация |
ИСТИНА ИНХС РАН |
||
A neurosteroid analogue photolabeling reagent labels the colchicines-binding site on tubulin: a mass spectrometric analysisстатья
Статья опубликована в высокорейтинговом журнале
Статья опубликована в журнале из списка Web of Science и/или Scopus- Авторы: Chen Z., Chen LH, Akentieva N., Lichti CF, Darbandi R., Hastings R., Lichti Ch F., Gross J., Covey D.F., Reichert D.E., Townsend R., Alex S.Evers
- Журнал: Electrophoresis
- Том: 33
- Номер: 4
- Год издания: 2011
- Издательство: John Wiley & Sons Ltd.
- Местоположение издательства: United Kingdom
- Первая страница: 666
- Последняя страница: 674
- Аннотация: Previous studies have shown that the neurosteroid analogue, 6-Azi-pregnanolone (6-AziP),photolabels voltage-dependent anion channels and proteins of approximately 55 kDa in rat brainmembranes. The present study used two dimensional electrophoresis and nano-electrosprayionization ion trap mass spectrometry (nano-ESI-MS) to identify the 55 kDa proteins (pI 4.8) asisoforms of β-tubulin. This identification was confirmed by immuno-blot and immunoprecipitation of photolabeled protein with anti-β-tubulin antibody and by the demonstration that 6-AziP photolabels purified bovine brain tubulin in a concentration-dependent pattern. To identify the photolabeling sites, purified bovine brain tubulin was photolabeled with 6-AziP, digested with trypsin, and analyzed by matrix-assisted laser desorption/ionization mass spectrometry (MALDI). A 6-AziP adduct of TAVCDIPPR (m/z=1287.77), a β-tubulin specific peptide, was detected by MALDI. High resolution LC-MS/MS analysis identified that 6-AziP was covalently bound to cysteine 354 (Cys-354), previously identified as a colchicine binding site. 6-AziP photolabeling was inhibited by 2-methoxyestradiol, an endogenous derivative of estradiol thought to bind to the colchicine site. Structural modeling predicted that neurosteroids could dock in this colchicine site at the interface between α- and β-tubulin with the photolabeling group of 6-AziP positioned proximate to Cys-354.
- Добавил в систему: Акентьева Наталья Павловна
Прикрепленные файлы
| № | Имя | Описание | Имя файла | Размер | Добавлен |
|---|---|---|---|---|---|
| 1. | Полный текст | Akentieva_and_Evers.pdf | 1,6 МБ | 8 июня 2022 [Akentieva] |