Отправить статью по E-mail Femtosecond and picosecond dynamics of recombinant bacteriorhodopsin primary reactions compared to the native protein in trimeric and monomeric forms
- Авторы: Smitienko O.1, Nekrasova O.2,3, Kudriavtsev A.1,3, Yakovleva M.1, Shelaev I.4, Gostev F.4, Dolgikh D.2,3,5, Kolchugina I.3, Nadtochenko V.4, Kirpichnikov M.2,3, Feldman T.1,3,5, Ostrovsky M.1,3
-
Учреждения:
- Emanuel Institute of Biochemical Physics
- Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry
- Lomonosov Moscow State University
- Semenov Institute of Chemical Physics
- Pirogov Russian National Research Medical University
- Выпуск: Том 82, № 4 (2017)
- Страницы: 490-500
- Раздел: Article
- URL: https://journals.rcsi.science/0006-2979/article/view/151351
- DOI: https://doi.org/10.1134/S0006297917040113
- ID: 151351
Цитировать
Открытый доступ
Доступ предоставлен
Только для подписчиков
Аннотация
Photochemical reaction dynamics of the primary events in recombinant bacteriorhodopsin (bRrec) was studied by femtosecond laser absorption spectroscopy with 25-fs time resolution. bRrec was produced in an Escherichia coli expression system. Since bRrec was prepared in a DMPC–CHAPS micelle system in the monomeric form, its comparison with trimeric and monomeric forms of the native bacteriorhodopsin (bRtrim and bRmon, respectively) was carried out. We found that bRrec intermediate I (excited state of bR) was formed in the range of 100 fs, as in the case of bRtrim and bRmon. Further processes, namely the decay of the excited state I and the formation of intermediates J and K of bRrec, occurred more slowly compared to bRtrim, but similarly to bRmon. The lifetime of intermediate I, judging from the signal of ΔAESA(470-480 nm), was 0.68 ps (78%) and 4.4 ps (22%) for bRrec, 0.52 ps (73%) and 1.7 ps (27%) for bRmon, and 0.45 ps (90%) and 1.75 ps (10%) for bRtrim. The formation time of intermediate K, judging from the signal of ΔAGSA(625-635 nm), was 13.5 ps for bRrec, 9.8 ps for bRmon, and 4.3 ps for bRtrim. In addition, there was a decrease in the photoreaction efficiency of bRrec and bRmon as seen by a decrease in absorbance in the differential spectrum of the intermediate K by ~14%. Since photochemical properties of bRrec are similar to those of the monomeric form of the native protein, bRrec and its mutants can be considered as a basis for further studies of the mechanism of bacteriorhodopsin functioning.
Об авторах
O. Smitienko
Emanuel Institute of Biochemical Physics
Автор, ответственный за переписку.
Email: djolia@gmail.com
Россия, Moscow, 119334
O. Nekrasova
Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry; Lomonosov Moscow State University
Email: djolia@gmail.com
Россия, Moscow, 117997; Moscow, 119991
A. Kudriavtsev
Emanuel Institute of Biochemical Physics; Lomonosov Moscow State University
Email: djolia@gmail.com
Россия, Moscow, 119334; Moscow, 119991
M. Yakovleva
Emanuel Institute of Biochemical Physics
Email: djolia@gmail.com
Россия, Moscow, 119334
I. Shelaev
Semenov Institute of Chemical Physics
Email: djolia@gmail.com
Россия, Moscow, 119991
F. Gostev
Semenov Institute of Chemical Physics
Email: djolia@gmail.com
Россия, Moscow, 119991
D. Dolgikh
Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry; Lomonosov Moscow State University; Pirogov Russian National Research Medical University
Email: djolia@gmail.com
Россия, Moscow, 117997; Moscow, 119991; Moscow, 1117997
I. Kolchugina
Lomonosov Moscow State University
Email: djolia@gmail.com
Россия, Moscow, 119991
V. Nadtochenko
Semenov Institute of Chemical Physics
Email: djolia@gmail.com
Россия, Moscow, 119991
M. Kirpichnikov
Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry; Lomonosov Moscow State University
Email: djolia@gmail.com
Россия, Moscow, 117997; Moscow, 119991
T. Feldman
Emanuel Institute of Biochemical Physics; Lomonosov Moscow State University; Pirogov Russian National Research Medical University
Email: djolia@gmail.com
Россия, Moscow, 119334; Moscow, 119991; Moscow, 1117997
M. Ostrovsky
Emanuel Institute of Biochemical Physics; Lomonosov Moscow State University
Email: djolia@gmail.com
Россия, Moscow, 119334; Moscow, 119991
