Hydrogen sulfide deactivates common nitrobenzofurazan-based fluorescent thiol labeling reagents.
Hydrogen sulfide deactivates common nitrobenzofurazan-based fluorescent thiol labeling reagents.
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DOI：
10.1021/ac501193r
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发表时间：
2014-06-17
影响因子：
7.4
通讯作者：
Pluth, Michael D.
中科院分区：
文献类型：
--
作者：
Montoya, Leticia A.;Pluth, Michael D.
硫醇和硫化氢(H_2S)等含硫化合物在生物结构和功能中起着重要但又不同的作用。分离硫醇和硫化氢的生物作用的一个主要挑战是开发工具来有效地分离这些含硫羟基化合物的反应性。为了应对这一挑战，我们报道了常见的亲电荧光硫醇标记剂，包括基于硝基苯并呋喃的支架、马来酰亚胺、烷化剂和亲电醛，对半胱氨酸和硫化氢的不同响应。虽然硫化氢与所有被研究的支架都发生了反应，但对每种支架的光物理响应都有显著的不同。以马来酰亚胺为基础的烷基化和缩醛硫醇标记剂在处理硫化氢时荧光反应减弱。相反，以硝基苯并呋喃为基础的标记试剂会被硫化氢失活。此外，在硫醇活化的硝基呋喃类试剂中加入H_2S后，荧光信号减弱，从而建立了硫醇标记试剂在H_2S存在下的不相容性。综上所述，这些研究强调了硫醇和硫化氢对常见硫醇标记试剂的不同反应能力，并建议在标记或测量产生硫化氢的细胞环境中的硫醇时必须足够小心，因为可能会出现假阳性和腐蚀反应。
Sulfhydryl-containing compounds, including thiols and hydrogen sulfide (H2S), play important but differential roles in biological structure and function. One major challenge in separating the biological roles of thiols and H2S is developing tools to effectively separate the reactivity of these sulfhydryl-containing compounds. To address this challenge, we report the differential responses of common electrophilic fluorescent thiol labeling reagents, including nitrobenzofurazan-based scaffolds, maleimides, alkylating agents, and electrophilic aldehydes, toward cysteine and H2S. Although H2S reacted with all of the investigated scaffolds, the photophysical response to each scaffold was significantly different. Maleimide-based, alkylating, and aldehydic thiol labeling reagents provided a diminished fluorescence response when treated with H2S. By contrast, nitrobenzofurazan-based labeling reagents were deactivated by H2S addition. Furthermore, the addition of H2S to thiol-activated nitrobenzofurazan-based reagents reduced the fluorescence signal, thus establishing the incompatibility of nitrobenzofurazan-based thiol labeling reagents in the presence of H2S. Taken together, these studies highlight the differential reactivity of thiols and H2S toward common thiol-labeling reagents and suggest that sufficient care must be taken when labeling or measuring thiols in cellular environments that produce H2S due to the potential for both false-positive and eroded responses.
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影响因子：
7.8
作者：
Lin, Vivian S.;Chang, Christopher J.
通讯作者：
Chang, Christopher J.
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影响因子：
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作者：
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通讯作者：
Snyder SH
影响因子：
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作者：
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通讯作者：
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