Non-heme iron protein: a potential target of nitric oxide in acute cardiac allograft rejection.
Non-heme iron protein: a potential target of nitric oxide in acute cardiac allograft rejection.
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非血红素铁蛋白：一氧化氮在急性心脏同种异体移植排斥反应中的潜在靶标。
DOI：
10.1073/pnas.0636938100
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发表时间：
2003
期刊：
影响因子：
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通讯作者：
Griffith,OwenW
中科院分区：
文献类型：
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作者：
Pieper,GalenM;Halligan,NadineLN;Hilton,Gail;Konorev,EugeneA;Felix,ChristopherC;Roza,AllanM;Adams,MarkB;Griffith,OwenW
在心脏移植急性排斥反应中，我们检测了非血红素蛋白的亚硝化铁和铁-S簇蛋白乌头酸酶的活性。供体心脏的异位移植是在组织相容性匹配(同种异体移植：lewis→lewis)和不相合(同种异体移植：wistar-Furth→lewis)大鼠中进行的。术后第4~6天，Western印迹分析和免疫组织化学显示同种异体移植物中有诱导型一氧化氮合酶(INOS)蛋白表达，而同种异体移植物中未见。EPR谱显示同种异体移植心脏和正常心脏的背景信号atg=2.003(半喹酮)和g=2.02和g=1.94(Fe-S簇蛋白)。相反，在POD4的同种异体移植物中，出现了一个新的轴向信号ATG=2.04和G=2.02，该信号归因于非血红素蛋白亚硝化形成的二亚硝基-铁复合体。这一信号出现在血红素蛋白显著亚硝化时或之前。非血红素蛋白的铁亚硝化与非亚硝化的铁-S簇信号ATG=1.94的减少是一致的。经POD_4处理后，乌头酸酶活性降至同基因对照的≈的50%。环孢素可阻断(1)血浆硝酸盐+亚硝酸盐的升高，(2)诱导型一氧化氮合酶蛋白的上调，(3)铁-S簇EPR信号的减少，(4)二亚硝基-铁络合物的形成，(5)乌头酸酶活性的丧失。受体用选择性诱导型一氧化氮合酶抑制剂L-N6-(1-亚氨基乙基)赖氨酸处理后，移植物内二亚硝基-铁复合体的形成和乌头酸酶活性的丧失也被抑制。本报告显示了一种重要的非血红素铁-S簇蛋白在急性固体器官移植排斥反应中的靶向性。
We examined iron nitrosylation of non-heme protein and enzymatic activity of the Fe-S cluster protein, aconitase, in acute cardiac allograft rejection. Heterotopic transplantation of donor hearts was performed in histocompatibility matched (isografts: Lewis → Lewis) and mismatched (allografts: Wistar–Furth → Lewis) rats. On postoperative days (POD) 4–6, Western blot analysis and immunohistochemistry revealed inducible nitric-oxide synthase (iNOS) protein in allografts but not isografts. EPR spectroscopy revealed background signals atg= 2.003 (for semiquinone) andg= 2.02 andg= 1.94 (for Fe-S cluster protein) in isografts and normal hearts. In contrast, in allografts on POD4, a new axial signal atg= 2.04 andg= 2.02 appeared that was attributed to the dinitrosyl–iron complex formed by nitrosylation of non-heme protein. Appearance of this signal occurred at or before significant nitrosylation of heme protein. Iron nitrosylation of non-heme protein was coincidental with decreases in the nonnitrosylated Fe-S cluster signal atg= 1.94. Aconitase enzyme activity was decreased to ≈50% of that observed in isograft controls by POD4. Treatment with cyclosporine blocked the (i) elevation of plasma nitrate + nitrite, (ii) up-regulation of iNOS protein, (iii) decrease in Fe-S cluster EPR signal, (iv) formation of dinitrosyl–iron complexes, and (v) loss of aconitase enzyme activity. Formation of dinitrosyl–iron complexes and loss of aconitase activity within allografts also was inhibited by treatment of recipients with a selective iNOS inhibitor,l-N6-(1-iminoethyl)lysine. This report shows targeting of an important non-heme Fe-S cluster protein in acute solid organ transplant rejection.