title: Light- and pH-dependent conformational changes in protein structure induce strong bending of purple membranes--active membranes studied by cryo-SEM.
abstract: Bacteriorhodopsin (BR) undergoes a conformational change during the photocycle and the proton transport through the membrane. For the first time, we could demonstrate by direct imaging of freely suspended native purple membranes (PMs) that the flat disk-like shape of PMs changes dramatically as soon as most of the BRs are in a state characterized by a deprotonated Schiff base. Light-induced shape changes are easily observed with mutated BRs of the BR-D96N type, i.e., all variants which show an increased M 2 lifetime. On the other hand, large-scale shape changes are induced by pH changes with PM containing mutated BRs of the BR-D85T type, where Asp85 is replaced for a neutral amino acid. In such PMs, all BRs are titrated simultaneously and the resulting shape of the membranes depends on the initial shape only. As the majority of PMs in the "flat" state are more or less round disks, the bent membranes often comprise bowl-like and tube-like bent structures. The method presented here enables one to derive size changes of membrane-embedded BRs on the single molecule level from "macroscopic", easily accessible data like the curvature radii observed in cryo-SEM. The potential of BR as a pH-controlled and/or light-controlled microscaled biological actuator needs further consideration.
authors: Department of Chemistry, University of Marburg, and Material Sciences Center, Hans-Meerwein-Strasse, Marburg, Germany.
authors: Daniel
authors: D
authors: Rhinow
authors: Norbert A
authors: NA
authors: Hampp
mesh_terms: D019943:Amino Acid Substitution; D001436:Bacteriorhodopsins / Q000737:chemistry* / Q000528:radiation effects; D006863:Hydrogen-Ion Concentration; D008027:Light; D008855:Microscopy, Electron, Scanning; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D018717:Purple Membrane / Q000378:metabolism / Q000648:ultrastructure*; D012545:Schiff Bases / Q000737:chemistry
chemical_list: D012545:Schiff Bases; D001436:Bacteriorhodopsins