Molecular orientation distributions in protein films .2. Site-directed immobilization of yeast cytochrome c on thiol-capped, self-assembled monolayers

被引:71
作者
Wood, LL [1 ]
Cheng, SS [1 ]
Edmiston, PL [1 ]
Saavedra, SS [1 ]
机构
[1] UNIV ARIZONA,DEPT CHEM,TUCSON,AZ 85721
关键词
D O I
10.1021/ja9623673
中图分类号
O6 [化学];
学科分类号
0703 ;
摘要
Molecular orientation in films of yeast cytochrome c immobilized via disulfide bonding between cysteine 102 and the thiol tail groups of self-assembled monolayers (SAMs) coated on planar 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 planar integrated optical waveguide-attenuated total reflection geometry, and emission anisotropy, measured in a total internal reflection fluorescence geometry. The mean heme tilt angle and angular distribution about the mean were recovered using a Gaussian model for the orientation distribution. These data are the first orientation distribution measurements reported for a protein film immobilized using a site-directed bonding strategy. The results show that the molecular architecture examined in this study does not produce a highly oriented protein film. A significant fraction of the immobilized cytochrome c is nonspecifically adsorbed to the SAM surface, which produces a relatively broad distribution of heme orientations.
引用
收藏
页码:571 / 576
页数:6
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