Molecular orientation distributions in protein films: III. Yeast cytochrome c immobilized on pyridyl disulfide-capped phospholipid bilayers.
Molecular orientation in a hydrated monolayer film of yeast cytochrome c, immobilized via disulfide bonding between Cys-102 and a pyridyl disulfide-capped phospholipid bilayer deposited from an air-water interface onto glass substrates, was investigated. The orientation distribution of the heme groups in the protein film was determined using a combination of absorption linear dichroism, measured in a planarintegrated optical waveguide-attenuated total reflection geometry- and fluorescence anisotropy, measured in a total internal reflection geometry. A gaussian model for the orientation distribution was used to recover the mean heme tilt angle and angular distribution about the mean, which were 40 and 11 degrees, respectively. Additional experiments showed that a large fraction of the cytochrome c was disulfide bonded to the bilayer, which correlates with the high degree of macroscopic order in the protein film. However, a subpopulation of yeast cytochrome c molecules in the film (approximately 30% of the total) appeared to be nonspecifically adsorbed. The orientation distribution of this subpopulation was found to be much broader than the specifically bound fraction.
Edmiston
P L
PL
Department of Chemistry, University of Arizona, Tucson 85721, USA.
Saavedra
S S
SS
D000327:Adsorption; D001665:Binding Sites; D003545:Cysteine; D003574:Cytochrome c Group / Q000737:chemistry*; D019281:Dimerization; D004229:Dithiothreitol; D008051:Lipid Bilayers; D008958:Models, Molecular; D008968:Molecular Conformation; D016011:Normal Distribution; D010084:Oxidation-Reduction; D010713:Phosphatidylcholines / Q000737:chemistry; D010714:Phosphatidylethanolamines / Q000737:chemistry*; D011312:Pressure; D012441:Saccharomyces cerevisiae
D003574:Cytochrome c Group; C112018:DOPE-PDS; D008051:Lipid Bilayers; D010713:Phosphatidylcholines; D010714:Phosphatidylethanolamines; C017251:1,2-oleoylphosphatidylcholine; D003545:Cysteine; D004229:Dithiothreitol