danДоклады Национальной академии наук БеларусиDoklady of the National Academy of Sciences of Belarus1561-83232524-2431The Republican Unitary Enterprise Publishing House "Belaruskaya Navuka"dan-405Research ArticleХИМИЯCHEMISTRYИССЛЕДОВАНИЕ ТРАНСМЕМБРАННОГО БЕЛКА CD79B МЕТОДОМ МНОГОМЕРНОЙ ИМПУЛЬСНОЙ ЯМР СПЕКТРОСКОПИИSTUDY OF THE TRANSMEMBRANE PROTEIN CD79B BY MULTIDIMENSIONAL PULSE NMR SPECTROSCOPYПанкратоваЕ. В.PankratovaE. V.

мл. науч. сотрудник

Junior researcher

pankratovaelena3@gmail.comБритиковВ. В.BritikovV. V.

науч. сотрудник

Researcher

vvbritikov@gmail.comУсановС. А.UsanovS. A.

член-корреспондент, д-р хим. наук, профессор

Corresponding Member, D. Sc. (Chemistry), Professor

usanov@iboch.bas-net.byИнститут биоорганической химии НАН Беларуси, МинскInstitute of Bioorganic Chemistry of the National Academy of Sciences of Belarus, Minsk2017290420176123950Copyright © Панкратова Е.В., Бритиков В.В., Усанов С.А., 20172017Панкратова Е.В., Бритиков В.В., Усанов С.А.Pankratova E.V., Britikov V.V., Usanov S.A.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://doklady.belnauka.by/jour/article/view/405

В настоящей работе с использованием системы бесклеточной экспрессии получены меченые стабильными изотопами углерода-13 и азота-15 белки CD79A/CD79B. Установлено, что целевые белки обнаруживаются в осадке бесклеточной системы экспрессии, что согласуется с их мембранной природой и наличием трансмембранного домена в их составе. Определены физико-химические параметры образцов CD79A/CD79B с целью получения многомерных корреляционных ЯМР спектров высокого разрешения. Анализ полученных корреляционных спектров свидетельствует, что CD79B при выбранных условиях находится в неупорядоченном состоянии. Расщепление сигнала от NH- группы боковой цепи единственного остатка триптофана указывает на наличие медленных конформационных превращений в этой области полипептидной цепи. Добавление трифторуксусной кислоты к раствору CD79B в диметил- сульфоксиде приводит к разрушению межмолекулярных связей «белковой мицеллы» и образованию его мономерной формы, хорошо детектируемой ЯМР спектроскопией.

 

In the present work, using the cell-free expression system, we prepared the proteins CD79A/CD79B labeled by stable isotopes of carbon-13 and nitrogen-15. It is shown that target proteins are mostly localized in the pellet of the cell-free expression system, which is consistent with their membrane nature and the presence of a transmembrane domain in their structure. Physicochemical parameters of the CD79A/CD79B samples were defined to obtain the multidimensional correlation NMR spectra of high resolution. The analysis of the obtained correlation spectra shows that under experimental conditions, CD79B exists in the disordered state. The splitting of the signal from the NH-group of the side chain of single tryptophan residue indicates the presence of slow conformational transitions in this region of the polypeptide chain. The addition of the trifluoroacetic acid to the solution of CD79B in DMSO leads to the destruction of the intermolecular bonds of “protein micelles” and the formation of its monomeric form that is well detectable by NMR.

 

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