Surface Coverage Dependent Renaturation of Oxidized Cytochrome c Adsorbed to a Fused Silica Surface

Surface Coverage Dependent Renaturation of Oxidized Cytochrome c Adsorbed to a Fused Silica Surface S. A. Mistry, S. A. Hocker, V. S. Fahrenbach, T. L. Benz, G. C. Campanello G. C. Hoops, T. A. Hopkins, M.-C. Su Department of Chemistry, Butler University, Indianapolis, IN 46208

Cytochrome c ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],Heme Tryptophan Tyrosine

Cyt c Function Voet, Donald; Voet, Judith; Pratt, Charlotte. Fundamentals of Biochemistry Upgrade Ed. 2001:501 Plays a major role in the electron transport chain in the inner membrane of mitochondria Shuttles electrons between complexes III & IV

Solution Absorption 20 µ M [Cyt c] in 10mM pH 4.7 Succinate Buffer Cox, M., Nelson, D. Principles of Biochemistry 2000:194 Soret Band ,[object Object],[object Object],[object Object],[object Object],[object Object]

Solution Alcohol Denatured ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]

ATR spectroscopy of cyt c on fused silica surface Cheng, Y.-Y.; Lin, S. H.; Chang, H.-C.; Su, M.-C.: Probing Adsorption, Orientation and Conformational Changes of Cytochrome c on Fused Silica Surfaces with the Soret Band. J. Phys. Chem. A pp. 10687, 107 (49) 2003 ,[object Object],[object Object],[object Object],Detector Light Source ,[object Object],[object Object],[object Object],[object Object],Prism θ Glass plate O-ring Sample solution To detector

Alcohol denatured Isotherm ,[object Object],[object Object],[object Object],[object Object]

Surface Spectra: ,[object Object],[object Object],[object Object],*Note the spectrum for 0.1µM is enlarged ,[object Object],[object Object],[object Object],[object Object],[object Object]

Surface adsorbed vs. Solution: [Succinate Buffer] = 10 mM pH = 4.7 [NaCl] = 150 mM *Note the spectrum for 0.1 µM is enlarged ,[object Object],[object Object],[object Object],[object Object],[object Object]

Surface adsorbed vs. Solution: [Succinate Buffer] = 10 mM 60% n-propanol [NaCl] = 150 mM pH = 4.7 ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]

Protein-Protein Interactions: ,[object Object],[object Object],[object Object]

Conclusion ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]

Special Thanks To: ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]

Surface Coverage Dependent Renaturation of Oxidized Cytochrome c Adsorbed to a Fused Silica Surface

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Surface Coverage Dependent Renaturation of Oxidized Cytochrome c Adsorbed to a Fused Silica Surface