Modification by homocysteine thiolactone affects redox status of cytochrome c

被引:60
作者
Perla-Kajan, Joanna
Marczak, Lukasz
Kajan, Laszlo
Skowronek, Pawel
Twardowski, Tomasz
Jakubowski, Hieronim
机构
[1] Polish Acad Sci, Inst Bioorgan Chem, PL-61704 Poznan, Poland
[2] Univ Agr, Dept Biochem & Biotechnol, PL-60637 Poznan, Poland
[3] BioInfoBank Inst, PL-60744 Poznan, Poland
[4] Adam Mickiewicz Univ, Dept Chem, PL-60780 Poznan, Poland
[5] Univ Med & Dent New Jersey, Dept Neurol & Mol Genet, New Jersey Med Sch, Newark, NJ 07101 USA
关键词
D O I
10.1021/bi602463m
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
Homocysteine (Hcy)-thiolactone mediates a post-translational incorporation of Hcy into protein in humans. Protein N-homocysteinylation is detrimental to protein structure and function and is linked to pathophysiology of hyperhomocysteinemia observed in humans and experimental animals. The modification by Hcy-thiolactone can be detrimental directly by affecting the function of an essential lysine residue or indirectly by interfering with the function of other essential residues or cofactors. Previous work has shown that cytochrome c is very sensitive to Hcy-thiolactone, which causes formation of N-Hcy-cytochrome c multimers. However, it was unclear what sites in cytochrome c were prone to Hcy attachment and whether N-linked Hcy can affect the structure and redox function of cytochrome c. Here we show that 4 lysine residues (Lys8 or -13, Lys86 or -87, Lys99, and Lys100) of cytochrome c are susceptible to N-homocysteinylation. We also show that N-homocysteinylation of 1 mol of lysine/mol of protein affects the redox state of the heme ligand of cytochrome c by rendering it reduced. The modification causes subtle structural changes, manifested as increased resistance of the N-Hcy-cytochrome c to proteolysis by trypsin, chymotrypsin, and Pronase. However, no major secondary structure perturbations were observed as shown by circular dichroism spectroscopy. Our data illustrate how N-homocysteinylation can interfere with the function of heme-containing proteins.
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页码:6225 / 6231
页数:7
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