title: Conformational toggling of yeast iso-1-cytochrome C in the oxidized and reduced states.
abstract: To convert cyt c into a peroxidase-like metalloenzyme, the P71H mutant was designed to introduce a distal histidine. Unexpectedly, its peroxidase activity was found even lower than that of the native, and that the axial ligation of heme iron was changed to His71/His18 in the oxidized state, while to Met80/His18 in the reduced state, characterized by UV-visible, circular dichroism, and resonance Raman spectroscopy. To further probe the functional importance of Pro71 in oxidation state dependent conformational changes occurred in cyt c, the solution structures of P71H mutant in both oxidation states were determined. The structures indicate that the half molecule of cyt c (aa 50-102) presents a kind of "zigzag riveting ruler" structure, residues at certain positions of this region such as Pro71, Lys73 can move a big distance by altering the tertiary structure while maintaining the secondary structures. This finding provides a molecular insight into conformational toggling in different oxidation states of cyt c that is principle significance to its biological functions in electron transfer and apoptosis. Structural analysis also reveals that Pro71 functions as a key hydrophobic patch in the folding of the polypeptide of the region (aa 50-102), to prevent heme pocket from the solvent.
authors.affiliation: State Key Laboratory of Bioorganic and Natural Products Chemistry, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, Shanghai, China.
authors.forename: Wenxian
authors.initials: W
authors.lastname: Lan
authors.forename: Zhonghua
authors.initials: Z
authors.lastname: Wang
authors.forename: Zhongzheng
authors.initials: Z
authors.lastname: Yang
authors.forename: Jing
authors.initials: J
authors.lastname: Zhu
authors.forename: Tianlei
authors.initials: T
authors.lastname: Ying
authors.forename: Xianwang
authors.initials: X
authors.lastname: Jiang
authors.forename: Xu
authors.initials: X
authors.lastname: Zhang
authors.forename: Houming
authors.initials: H
authors.lastname: Wu
authors.forename: Maili
authors.initials: M
authors.lastname: Liu
authors.forename: Xiangshi
authors.initials: X
authors.lastname: Tan
authors.forename: Chunyang
authors.initials: C
authors.lastname: Cao
authors.forename: Zhong-Xian
authors.initials: ZX
authors.lastname: Huang
mesh_terms: D019943:Amino Acid Substitution; D017209:Apoptosis; D001665:Binding Sites; D003575:Cytochromes c1 / Q000737:chemistry* / Q000235:genetics / Q000502:physiology*; D004579:Electron Transport; D005656:Fungal Proteins / Q000737:chemistry*; D057927:Hydrophobic and Hydrophilic Interactions; D007501:Iron; D010084:Oxidation-Reduction; D011487:Protein Conformation; D013057:Spectrum Analysis
chemical_list: D005656:Fungal Proteins; D003575:Cytochromes c1; D007501:Iron
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