[1H-15N NMR signal assignment and secondary structure of bacteriorhodopsin (1-231) in solution].
15N-Labeled de-(232-248)-bacteriorhodopsin [BR(1-231)] was solubilized in 1:1 chloroform-methanol solvent mixture that contained 1.0 M 2HCO2N2H4 and mimic membrane medium. Resonances in the 1H-15N heteronuclear multiple-quantum coherence (HMQC) spectrum of BR (1-231) were assigned using the data of two- and three-dimensional NMR experiments. Of 117 cross-peaks present in the 1H-15N HMQC spectrum, 98 were assigned to residues in 1-75 and 193-231 segments of the protein. Almost all cross-peaks that correspond to the 76-192 segment were absent in the HMQC spectrum (except for six cross-peaks from the side chains and 14 cross-peaks from the backbone). Deuterium exchange rates of amide protons and cross-peaks of nuclear Overhauser effect helped to localize helices A (residues 8-30), B (residues 40-65), and G (residues 198-226). The periodicity in the rates of deuterium exchange of NH protons of helices A, B, and G was explained by the compact arrangement of these helices in the protein globule. The broadening of signals from six residues in helix G, which, according to the electron cryomicroscopy model of bacteriorhodopsin, is in contact with the NMR-unobservable bundle of helices CDEF, indicates specific interactions of the helices in BR(1-231). These data suggest that BR(1-231) solubilized in an organic medium has a spatial structure similar to that in the electron cryomicroscopy model of BR.
Grabchuk
I A
IA
Orekhov
V Iu
VIu
Musina
L Iu
LIu
Arsen'ev
A S
AS
D000596:Amino Acids / Q000737:chemistry*; D001436:Bacteriorhodopsins / Q000737:chemistry* / Q000648:ultrastructure; D002725:Chloroform / Q000737:chemistry; D003903:Deuterium; D009682:Magnetic Resonance Spectroscopy; D000432:Methanol / Q000737:chemistry; D008854:Microscopy, Electron; D017433:Protein Structure, Secondary; D012996:Solutions
D000596:Amino Acids; D012996:Solutions; D001436:Bacteriorhodopsins; D002725:Chloroform; D003903:Deuterium; D000432:Methanol
Otnesenie 1H-15N signalov i vtorichnaya struktura Des-(232-248) bacteriorodopsina v rastvore.