Spectroscopic studies of bacteriorhodopsin fragments dissolved in organic solution.
Fourier transform infrared and UV fourth-derivative spectroscopies were used to study the secondary structure of bacteriorhodopsin and its chymotryptic and one of the sodium borohydride fragments dissolved in chloroform-methanol (1:1, v/v), 0.1 M LiClO4. The C1 fragment (helices C, D, E, F, and G) showed an alpha-helical content of about 53%, whereas C2 (helices A and B) had about 60%, and B2 (helices F and G) about 65% alpha-helix. The infrared main band indicated differences in alpha-helical properties between these fragments. These techniques were also used to obtain information on the interactions among helices. According to the results obtained from the hydrogen/deuterium exchange kinetics, about 40% of the amide protons of C2 are particularly protected against exchange, whereas for the C1 fragment this process is unexpectedly fast. UV fourth-derivative spectra of these samples were used to obtain information about the environment of Trp side chains. The results showed that the Trp residues of C2 are more shielded from the solvent than those of C1 or B2. The results of this work indicate that the specific interactions existing between the transmembrane segments induce different types of helical conformations in native bacteriorhodopsin.
Torres
J
J
Departament de Bioquímica i de Biologia Molecular, Facultat de Medicina, Universitat Autònoma de Barcelona, Spain.
Padrós
E
E
D001436:Bacteriorhodopsins / Q000737:chemistry* / Q000302:isolation & purification; D001894:Borohydrides; D002725:Chloroform; D002918:Chymotrypsin; D003903:Deuterium; D006217:Halobacterium / Q000378:metabolism; D006859:Hydrogen; D018020:Lithium Compounds; D000432:Methanol; D008958:Models, Molecular; D010446:Peptide Fragments / Q000737:chemistry* / Q000302:isolation & purification; D010472:Perchlorates; D017433:Protein Structure, Secondary; D012996:Solutions; D013056:Spectrophotometry, Ultraviolet; D017550:Spectroscopy, Fourier Transform Infrared
D001894:Borohydrides; D018020:Lithium Compounds; D010446:Peptide Fragments; D010472:Perchlorates; D012996:Solutions; D001436:Bacteriorhodopsins; D002725:Chloroform; D006859:Hydrogen; C025364:sodium borohydride; D003903:Deuterium; D002918:Chymotrypsin; C054684:lithium perchlorate; D000432:Methanol