In Situ Labeling and Distance Measurements of Membrane Proteins in E. coli Using Finland and OX063 Trityl Labels

In Situ Labeling and Distance Measurements of Membrane Proteins in E. coli Using Finland and OX063 Trityl Labels Научная публикация

Журнал

Chemistry - A European Journal
ISSN: 0947-6539 , E-ISSN: 1521-3765

Вых. Данные

Год: 2021, Том: 27, Номер: 7, Страницы: 2299-2304 Страниц : 6 DOI: 10.1002/chem.202004606

Ключевые слова

DEER or PELDOR; FTAM and OX063; membrane protein; spin labeling; beta-barrel assembly machinery (BAM) complex

Авторы

Ketter Sophie ¹ , Gopinath Aathira ¹ , Rogozhnikova Olga ² , Trukhin Dmitrii ² , Tormyshev Victor M. ² , Bagryanskaya Elena G. ² , Joseph Benesh ¹

Организации

1	(Данные Web of science) Goethe Univ Frankfurt, Inst Biophys, Dept Phys, Max von Laue Str 1, D-60438 Frankfurt, Germany
2	(Данные Web of science) RAS, SB, NN Vorozhtsov Novosibirsk Inst Organ Chem, Pr Lavrentieva 9, Novosibirsk 630090, Russia

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 beta-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 (T-M) of approximate to 5 mu s. All the trityls enabled distance measurements between BtuB and an orthogonally labeled substrate with high selectivity and sensitivity down to a few mu m concentration. Our data corroborate the BtuB and BamA conformations in the cellular environment of E. coli.

Ketter S. , Gopinath A. , Rogozhnikova O. , Trukhin D. , Tormyshev V.M. , Bagryanskaya E.G. , Joseph B.
In Situ Labeling and Distance Measurements of Membrane Proteins in E. coli Using Finland and OX063 Trityl Labels

Chemistry - A European Journal. 2021. V.27. N7. P.2299-2304. DOI: 10.1002/chem.202004606 WOS Scopus РИНЦ

Полный текст от издателя

Опубликована online:	14 янв. 2021 г.
Опубликована в печати:	1 февр. 2021 г.

Web of science	WOS:000607578700001
Scopus	2-s2.0-85099378059
РИНЦ	45006810
OpenAlex	W3101656168