The retinylidene chromophore mutant (Y185F) of bacteriorhodopsin, in which Tyr-185 is substituted by phenylalanine, is examined and compared with wild-type bacteriorhodopsin expressed in Escherichia coli; both were reinstituted similarly in vesicles. The Y185F mutant shows (at least) two distinct spectra at neutral pH. Upon light absorption, the blue species (which absorbs in the red) behaves a...

The retinal-protein complex bacteriorhodopsin mediates lightxnergy conversion in halobacteria. This chromoprotein is embedded in the cell membrane and forms insular regions. These can be isolated by membrane-fractionation procedures, and they consist of a lipid matrix containing bacteriorhodopsin molecules in a hexagonal crystalline arrangement called the purple membrane. Light-absorption by ba...

The function of three types of bacteriorhodopsins was compared: the wild-type, the bleached and retinal reconstituted and retinal deficient bacteriorhodopsin after retinal addition. The apparent pK(a) of the proton acceptor group for the bleached BR and retinal deficient BR shifted toward higher pH values compared to the wild-type BR. Fitting the photocycle model to the absorption kinetic signa...

The purple membrane is a two-dimensional crystalline lattice formed by bacteriorhodopsin and lipid molecules in the cytoplasmic membrane of Halobacterium salinarum. High-resolution structural studies, in conjunction with detailed knowledge of the lipid composition, make the purple membrane one of the best models for elucidating the forces that are responsible for the assembly and stability of i...

Homogenous bacteriorhodopsin-DPPC proteoliposomes were characterized and used as a target for the general anesthetic Enflurane. Dose–response curves for the protein and the lipids, in the very same sample, at high (2800) and low (360) lipid: bacteriorhodopsin molar ratios, were compared simultaneously. Above the main phase transition temperature of DPPC, there is no difference between the prote...

We theoretically and experimentally analyze the formation of thick Purple Membrane (PM) polyacrylamide (PA) films by means of optical spectroscopy by considering the absorption of bacteriorhodopsin and scattering. We have applied semiclassical quantum mechanical techniques for the calculation of absorption spectra by taking into account the Fano effects on the ground state of bacteriorhodopsin....

Bacteriorhodopsin is a rhodopsin-like protein found in the cell membrane of Halobacterium halobium. It shows an absorption maximum at 570 nm and, in the light, undergoes cyclic spectral changes which include a relatively long-lived complex absorbing maximally at 412 nm. Excitation profiles have been obtained with several laser frequencies for two vibrations in the resonance Raman spectrum of ba...

The folding mechanisms of helical membrane proteins remain largely uncharted. Here we characterize the kinetics of bacteriorhodopsin folding and employ φ-value analysis to explore the folding transition state. First, we developed and confirmed a kinetic model that allowed us to assess the rate of folding from SDS-denatured bacteriorhodopsin (bRU) and provides accurate thermodynamic information ...

The regeneration method of Khorana [J. Biol. Chem. 262 (1987) 9271] has been modified in order to study the effect of endogenous archaeabacterial lipids and, in particular, of glycocardiolipin (GlyC) in the refolding and chromophore regeneration of bacteriorhodopsin (BR). BR refolding and chromophore regeneration could be obtained in the presence of endogenous lipid mixtures containing or not c...

We have observed the spontaneous formation of transverse spatial patterns in a thin film of bacteriorhodopsin with a feedback mirror. Bacteriorhodopsin has a mixed absorptive-dispersive nonlinearity at the wavelength used in the experiments (633 nm). Threshold values of the incident intensity for observation of pattern formation are found from a linear stability analysis of a model that describ...