Characterization of cytochrome c free radical reactions with peptides by mass spectrometry

被引:49
作者
Deterding, LJ
Barr, DP
Mason, RP
Tomer, KB
机构
[1] NIEHS, Struct Biol Lab, Res Triangle Pk, NC 27709 USA
[2] NIEHS, Lab Pharmacol & Chem, Res Triangle Pk, NC 27709 USA
关键词
D O I
10.1074/jbc.273.21.12863
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The reactions of horse heart cytochrome c, hydrogen peroxide, and the spin trap 3,5-dibromo-4-nitrosobenzenesulfonic acid with a series of polypeptides were investigated using mass spectrometry. The mass spectra obtained from these reactions revealed that after a free radical has been generated on the heme containing protein horse heart cytochrome c, it can be transferred to other biomolecules. Zn addition, the number of free radicals transferred to the target molecule could be determined. Recipient peptides/proteins that contained a tyrosine and/or tryptophan amino acid residue were most susceptible to free radical transfer. Using tandem mass spectrometry, the location of the 3,5-dibromo-4-nitrosobenzenesulfonic acid radical adduct on the nonapeptide RWIILGLNK was unequivocally determined to be at the tryptophan residue. We also demonstrated that the presence of an antioxidant in the reaction mixture not only inhibits free radical formation on horse heart cytochrome c, but also interferes with the transfer of the free radical, once it has been formed on cytochrome c.
引用
收藏
页码:12863 / 12869
页数:7
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