A 1H NMR study of structurally relevant inter-segmental hydrogen bond in cytochrome c.
NMR signal arising from His 26 N(epsilon)H proton in horse and tuna ferrocytochromes c has been assigned. This His residue is highly conserved in most mitochondrial cytochromes c and X-ray crystallographic studies strongly suggested that its side-chain imidazole participates in an internal hydrogen bond network which is relevant to the stability of the non-helical protein folding near the heme active site. The shift and line width of the assigned signal indicated that this NH hydrogen is indeed involved in an internal hydrogen bond. On the basis of the X-ray crystal structures, the carbonyl oxygen of the residue at 44 is thought to act as a proton-acceptor for this hydrogen. The observation of nuclear Overhauser effect correlation between His 26 C(epsilon)H and Asn 31 main-chain amide NH proton signals in the present proteins also demonstrated the formation of the hydrogen bond between these residues. Consequently, the presence of a unique triad hydrogen bond network in these cytochromes c in solution has been confirmed. Taking advantage of the sensitivity of His 26 N(epsilon)H proton signal to the structural properties of this hydrogen bond network, influences of the presence of high concentration of salt or various concentrations of denaturant on the protein folding were inferred from the analysis of the NMR spectral parameters of the signal.
Yamamoto
Y
Y
Department of Chemistry, University of Tsukuba, Japan. yash@sakura.cc.tsukuba.ac.jp
D000595:Amino Acid Sequence; D000818:Animals; D003574:Cytochrome c Group / Q000737:chemistry*; D019791:Guanidine; D006639:Histidine / Q000737:chemistry / Q000378:metabolism; D006736:Horses; D006860:Hydrogen Bonding; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D009206:Myocardium / Q000737:chemistry; D019906:Nuclear Magnetic Resonance, Biomolecular; D009994:Osmolar Concentration; D011487:Protein Conformation; D011489:Protein Denaturation; D017510:Protein Folding; D014413:Tuna
D003574:Cytochrome c Group; D006639:Histidine; D019791:Guanidine