nioch.ru

Федеральное государственное бюджетное учреждение науки
Новосибирский институт органической химии им. Н.Н. Ворожцова
Сибирского отделения Российской академии наук

На сайте журнала Chemistry - A European Journal   (IF=4,857) опубликована статья  с участием сотрудников Института к.х.н.  О. Рогожниковой, к.х.н. Д. Трухина, к.х.н. В.М. Тормышева, д.ф.-м.н., проф. Е.Г.  Багрянской

 


In situ labeling and distance measurements of membrane proteins in E coli using Finland and OX063 trityl labels

Benesh Joseph, Sophie Ketter, Aathira Gopinath, Olga Rogozhnikova, Dmitrii Trukhin, Victor M. Tormyshev, Elena G. Bagryanskaya



Chemistry - A European Journal, 
First published 16 Novemberr 2020



 https://doi.org/10.1002/chem.202004606


Abstract

In situ investigation of membrane proteins is a challenging task. Previously we demonstrated that nitroxide labels combined with pulsed ESR spectroscopy is a promising tool for this purpose. However, the nitroxide labels suffer from poor stability, high background labeling, and low sensitivity. Here we show that Finland (FTAM) and OX063 based labels enable labeling of the cobalamin transporter BtuB and BamA, the central component of the β‐barrel assembly machinery (BAM) complex, in E coli. Compared to the methanethiosulfonate spin label (MTSL), trityl labels eliminated the background signals and enabled specific in situ labeling of the proteins with high efficiency. The OX063 labels show a long phase memory time (TM) of ~5 μs. All the trityls enabled distance measurements between BtuB and an orthogonally labeled substrate with high selectivity and sensitivity down to a few μM concentration. Our data corroborate the BtuB and BamA conformations in the cellular environment of E. coli.

Альметрики: 


Метрики PlumX теперь доступны в Scopus: узнайте, как другие ученые используют ваши исследования