A spectroscopic study of uranyl-cytochrome b5/cytochrome c interactions.
Uranium is harmful to human health due to its radiation damage and the ability of uranyl ion (UO2(2+)) to interact with various proteins and disturb their biological functions. Cytochrome b5 (cyt b5) is a highly negatively charged heme protein and plays a key role in mediating cytochrome c (cyt c) signaling in apoptosis by forming a dynamic cyt b5-cyt c complex. In previous molecular modeling study in combination with UV-Vis studies, we found that UO2(2+) is capable of binding to cyt b5 at surface residues, Glu37 and Glu43. In this study, we further investigated the structural consequences of cyt b5 and cyt c, as well as cyt b5-cyt c complex, upon uranyl binding, by fluorescence spectroscopic and circular dichroism techniques. Moreover, we proposed a uranyl binding site for cyt c at surface residues, Glu66 and Glu69, by performing a molecular modeling study. It was shown that uranyl binds to cyt b5 (KD=10 μM), cyt c (KD=87 μM), and cyt b5-cyt c complex (KD=30 μM) with a different affinity, which slightly alters the protein conformation and disturbs the interaction of cyt b5-cyt c complex. Additionally, we investigated the functional consequences of uranyl binding to the protein surface, which decreases the inherent peroxidase activity of cyt c. The information of uranyl-cyt b5/cyt c interactions gained in this study likely provides a clue for the mechanism of uranyl toxicity.
School of Chemistry and Chemical Engineering, University of South China, Hengyang 421001, China.
Mei-Hui
MH
Sun
Shuang-Quan
SQ
Liu
Ke-Jie
KJ
Du
Chang-Ming
CM
Nie
Ying-Wu
YW
Lin
D000818:Animals; D002417:Cattle; D015786:Cytochromes b5 / Q000737:chemistry / Q000378:metabolism*; D045304:Cytochromes c / Q000737:chemistry / Q000378:metabolism*; D006736:Horses; D006801:Humans; D007477:Ions; D007700:Kinetics; D008958:Models, Molecular; D009195:Peroxidase / Q000378:metabolism; D011485:Protein Binding; D013050:Spectrometry, Fluorescence; D014501:Uranium / Q000737:chemistry / Q000378:metabolism*
D007477:Ions; D014501:Uranium; D045304:Cytochromes c; D015786:Cytochromes b5; D009195:Peroxidase
Fluorescence; Heme protein; Peroxidase; Protein–protein interaction; Uranium