Effects of Triton X-100 on proton transfer and in the photocycle of archaerhodopsin 4.
Archaerhodopsin 4 (AR4), a retinal-containing membrane protein, exhibits a reversed order of proton release and uptake at neutral pH, as compared to the well-known bacteriorhodopsin (BR). In a preceding report, we stated that Triton X-100 solubilized the claret membrane containing AR4 (CM) into monomeric proteins and altered the time order in AR4 at neutral pH. The present study examined the mechanism underlying this phenomenon. We employed a photoelectrochemical cell suitable for observation of the proton pumping behaviors of both the membrane patch and detergent-solubilized proteins over a wide pH range. The pK(a) values of the proton release complex (PRC) in the initial state and the M state were determined with this device. The pK(a) of PRC of monomeric AR4 decreased to a value lower than 7.0 in the photocycle, allowing early proton release at neutral pH. The pK(a) of PRC in the initial state was also strongly affected by solubilization.
State Key Laboratory of Molecular Engineering of Polymers, Department of Macromolecular Science, Laboratory of Advanced Materials, Fudan University, Shanghai, China.
Yazhuo
Y
Wang
Jia
J
Wu
Ming
M
Ming
Yingchun
Y
Zhao
Jiandong
J
Ding
D019843:Archaeal Proteins / Q000502:physiology*; D006863:Hydrogen-Ion Concentration; D008565:Membrane Proteins; D017830:Octoxynol / Q000737:chemistry*; D017494:Proton Pumps / Q000737:chemistry*; D011522:Protons
D019843:Archaeal Proteins; D008565:Membrane Proteins; D017494:Proton Pumps; D011522:Protons; C055476:archaerhodopsin protein, Archaea; D017830:Octoxynol