title: Changes in global stability and local structure of cytochrome c upon substituting phenylalanine-82 with tyrosine.
abstract: We have examined the F82Y;C102T variant of Saccharomyces cerevisiae iso-1-cytochrome c using high-resolution proton nuclear magnetic resonance spectroscopy, chemical denaturation, and differential scanning calorimetry. Comparison of proton chemical shifts, paramagnetic shifts, and nuclear Overhauser effects indicates structural changes are localized to the vicinity of position 82. One alteration involves the rearrangement of the side chain of leucine-85. Using many more proton assignments than were available in the initial report [G. J. Pielak, R. A. Atkinson, J. Boyd, and R. J. P. Williams, Eur. J. Biochem. 177, 179-185 (1988)], a second alteration involving an interaction between arginine-13 and tyrosine-82 is observed. The interaction appears to involve a hydrogen bond with the eta-protons of arginine's guanido group acting as donor and tyrosine's phenolic eta-oxygen as acceptor. In spite of this potentially-stabilizing interaction, the free energy of denaturation decreases by approximately 2.4 kcal mol-1. Results are discussed with respect to alterations in the native and denatured states.
authors.affiliation: Department of Chemistry, University of North Carolina, Chapel Hill 27599-3290.
authors.forename: R M
authors.initials: RM
authors.lastname: Greene
authors.forename: S F
authors.initials: SF
authors.lastname: Betz
authors.forename: S
authors.initials: S
authors.lastname: Hilgen-Willis
authors.forename: D S
authors.initials: DS
authors.lastname: Auld
authors.forename: J B
authors.initials: JB
authors.lastname: Fencl
authors.forename: G J
authors.initials: GJ
authors.lastname: Pielak
mesh_terms: D003574:Cytochrome c Group / Q000737:chemistry*; D004795:Enzyme Stability; D015394:Molecular Structure; D010649:Phenylalanine / Q000737:chemistry*; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D014443:Tyrosine / Q000737:chemistry*
chemical_list: D003574:Cytochrome c Group; D014443:Tyrosine; D010649:Phenylalanine