Vectorially oriented monolayers of the cytochrome c/cytochrome oxidase bimolecular complex.
Vectorially oriented monolayers of yeast cytochrome c and its bimolecular complex with bovine heart cytochrome c oxidase have been formed by self-assembly from solution. Both quartz and Ge/Si multilayer substrates were chemical vapor deposited with an amine-terminated alkylsiloxane monolayer that was then reacted with a hetero-bifunctional cross-linking reagent, and the resulting maleimide endgroup surface then provided for covalent interactions with the naturally occurring single surface cysteine 102 of the yeast cytochrome c. The bimolecular complex was formed by further incubating these cytochrome c monolayers in detergent-solubilized cytochrome oxidase. The sequential formation of such monolayers and the vectorially oriented nature of the cytochrome oxidase was studied via meridional x-ray diffraction, which directly provided electron density profiles of the protein(s) along the axis normal to the substrate plane. The nature of these profiles is consistent with previous work performed on vectorially oriented monolayers of either cytochrome c or cytochrome oxidase alone. Furthermore, optical spectroscopy has indicated that the rate of binding of cytochrome oxidase to the cytochrome c monolayer is an order of magnitude faster than the binding of cytochrome oxidase to an amine-terminated surface that was meant to mimic the ring of lysine residues around the heme edge of cytochrome c, which are known to be involved in the binding of this protein to cytochrome oxidase.
Edwards
A M
AM
Department of Chemistry, University of Pennsylvania, Philadelphia 19104-6323, USA. edwards@jkb3.chem.upenn.edu
Blasie
J K
JK
Bean
J C
JC
D000465:Algorithms; D000818:Animals; D002417:Cattle; D003574:Cytochrome c Group / Q000737:chemistry* / Q000378:metabolism; D003576:Electron Transport Complex IV / Q000737:chemistry* / Q000378:metabolism; D005857:Germanium; D008929:Mitochondria, Heart / Q000201:enzymology; D011487:Protein Conformation; D011791:Quartz; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D012822:Silicon Dioxide; D013053:Spectrophotometry; D014961:X-Ray Diffraction
D003574:Cytochrome c Group; D005857:Germanium; D011791:Quartz; D012822:Silicon Dioxide; D003576:Electron Transport Complex IV