Protein adducts as prospective biomarkers of nevirapine toxicity.
Protein adducts as prospective biomarkers of nevirapine toxicity.
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DOI：
10.1021/tx100186t
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发表时间：
2010-11-15
影响因子：
4.1
通讯作者：
Marques MM
中科院分区：
文献类型：
--
作者：
Antunes AM;Godinho AL;Martins IL;Oliveira MC;Gomes RA;Coelho AV;Beland FA;Marques MM
奈韦拉平（NVP）是一种非核苷类逆转录酶抑制剂，用于抗人类免疫缺陷病毒1型（HIV-1），主要用于预防发展中国家的母婴HIV-1传播。尽管其临床疗效，NVP给药与各种毒性反应相关，包括肝毒性和皮疹。虽然NVP给药的不良反应的原因尚不清楚，但越来越多的证据支持代谢活化参与反应性亲电体。特别是，认为NVP代谢物12-羟基-NVP的II期活化介导NVP与生物亲核体的结合，这可能发生在毒性开始时。在本研究中，我们研究了人血清白蛋白和人血红蛋白中产生的共价加合物的性质和具体位置，该加合物通过与合成模型亲电体12-mesoxy-NVP（用作II期代谢物12-sulfoxy-NVP的替代物）在体外反应产生。通过两种不同的基于质谱的方法，液相色谱-电喷雾电离串联质谱（LC-ESI-MS/MS）和基质辅助激光解吸电离串联质谱（MALDI-TOF-TOF-MS），鉴定了多个修饰位点。这两种不同的方法，在某些情况下，提供了互补的信息，允许识别多个加合物，涉及半胱氨酸，赖氨酸，色氨酸，组氨酸，丝氨酸，和N-末端缬氨酸的血红蛋白。色氨酸不是共价蛋白质修饰的常见位点，其是在两种蛋白质中检测到的NVP修饰的氨基酸残基，并且通过LC-ESI-MS/MS和MALDI-TOF-TOF-MS两者一致地鉴定。色氨酸与NVP衍生的亲电试剂反应的倾向通过人血清白蛋白具有单个色氨酸残基的事实进一步强调，这表明了可用于生物监测目的的显著选择性。同样，NVP与血红蛋白末端缬氨酸的加合物，在N-烷基Edman降解后通过LC-ESI-MS/MS检测，似乎是NVP与蛋白质结合的容易评估的标志物。我们的研究结果证明了两种基于MS的方法用于表征NVP蛋白结合的优点和互补性，并提出了一系列合理的NVP毒性生物标志物，这些生物标志物应可用于监测NVP给药患者的毒性效应。
Nevirapine (NVP) is a non-nucleoside reverse transcriptase inhibitor used against the human immunodeficiency virus type-1 (HIV-1), mostly to prevent mother-to-child HIV-1 transmission in developing countries. Despite its clinical efficacy, NVP administration is associated with a variety of toxic responses that include hepatotoxicity and skin rash. Although the reasons for the adverse effects of NVP administration are still unclear, increasing evidence supports the involvement of metabolic activation to reactive electrophiles. In particular, Phase II activation of the NVP metabolite 12-hydroxy-NVP is thought to mediate NVP binding to bionucleophiles, which may be at the onset of toxicity. In the present study, we investigated the nature and specific locations of the covalent adducts produced in human serum albumin and human hemoglobin by reaction in vitro with the synthetic model electrophile 12-mesyloxy-NVP, used as a surrogate for the Phase II metabolite 12-sulfoxy-NVP. Multiple sites of modification were identified by two different mass spectrometry-based methodologies, liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) and matrix-assisted laser desorption ionization tandem mass spectrometry (MALDI-TOF-TOF-MS). These two distinct methodologies, which in some instances afforded complementary information, allowed the identification of multiple adducts involving cysteine, lysine, tryptophan, histidine, serine, and the N-terminal valine of hemoglobin. Tryptophan, which is not a common site of covalent protein modification, was the NVP-modified amino acid residue detected in the two proteins and consistently identified by both LC-ESI-MS/MS and MALDI-TOF-TOF-MS. The propensity of tryptophan to react with the NVP-derived electrophile is further emphasized by the fact that human serum albumin possesses a single tryptophan residue, which suggests a remarkable selectivity that may be useful for biomonitoring purposes. Likewise, the NVP adduct with the terminal valine of hemoglobin, detected by LC-ESI-MS/MS after N-alkyl Edman degradation, appears as an easily assessed marker of NVP binding to proteins. Our results demonstrate the merits and complementarity of the two MS-based methodologies for the characterization of protein binding by NVP and suggest a series of plausible biomarkers of NVP toxicity, that should be useful in the monitoring of toxicity effects in patients administered NVP.
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影响因子：
168.9
作者：
Jackson, JB;Musoke, P;Mmiro, F
通讯作者：
Mmiro, F
影响因子：
158.5
作者：
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Thaineua, V
DOI：
10.1016/j.ijheh.2007.01.024
发表时间：
2007-05-01
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影响因子：
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作者：
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通讯作者：
Jackson, JB