感应一氧化碳释放分子(CORM)不等同于感应一氧化碳:二(五氟苯基)五氮杂菲和 CORM-3 的情况。
Sensing a CO-Releasing Molecule (CORM) Does Not Equate to Sensing CO: The Case of DPHP and CORM-3.
机构信息
Department of Chemistry and Center for Diagnostics and Therapeutics, Georgia State University, Atlanta, Georgia 30303, United States.
出版信息
Anal Chem. 2023 Jun 13;95(23):9083-9089. doi: 10.1021/acs.analchem.3c01495. Epub 2023 Jun 1.
Carbon monoxide (CO) is an endogenous signaling molecule with demonstrated pharmacological effects. For studying CO biology, there is a need for sensitive and selective fluorescent probes for CO as research tools. In developing such probes, CO gas and/or commercially available metal-carbonyl-based "CO-releasing molecules" (CORMs) have been used as CO sources. However, new findings are steadily emerging that some of these commonly used CORMs do not release CO reliably in buffers commonly used for studying such CO probes and have very pronounced chemical reactivities of their own, which could lead to the erroneous identification of "CO probes" that merely detect the CORM used, not CO. This is especially true when the CO-sensing mechanism relies on chemistry that is not firmly established otherwise. Cu can quench the fluorescence of an imine-based fluorophore, DPHP, presumably through complexation. The Cu-quenched fluorescence was restored through the addition of CORM-3, a Ru-based CORM. This approach was reported as a new "strategy for detecting carbon monoxide" with the proposed mechanism being dependent on CO reduction of Cu to Cu under near-physiological conditions ( 2022, 94, 11298-11306). The study only used CORM-3 as the source of CO. CORM-3 has been reported to have very pronounced redox reactivity and is known not to release CO in an aqueous solution unless in the presence of a strong nucleophile. To assess whether the fluorescent response of the DPHP-Cu(II) cocktail to CORM-3 was truly through detecting CO, we report experiments using both pure CO and CORM-3. We confirm the reported DPHP-Cu(II) response to CORM-3 but not pure CO gas. Further, we did not observe the stated selectivity of DPHP for CO over sulfide species. Along this line, we also found that a reducing agent such as ascorbate was able to induce the same fluorescent turn-on as CORM-3 did. As such, the DPHP-Cu(II) system is not a CO probe and cannot be used to study CO biology. Corollary to this finding, it is critical that future work in developing CO probes uses more than a chemically reactive "CO donor" as the CO source. Especially important will be to confirm the ability of the "CO probe" to detect CO using pure CO gas or another source of CO.
一氧化碳(CO)是一种内源性信号分子,具有已证明的药理学作用。为了研究 CO 的生物学特性,需要使用敏感且选择性的荧光探针作为 CO 的研究工具。在开发这些探针时,CO 气体和/或市售的金属羰基基“CO 释放分子”(CORM)已被用作 CO 源。然而,新的发现不断涌现,表明这些常用的 CORM 中的一些在用于研究此类 CO 探针的常用缓冲液中不能可靠地释放 CO,并且具有非常明显的自身化学反应性,这可能导致错误地识别出“仅仅检测到所用 CORM 的 CO 探针”。当 CO 感应机制依赖于其他方式尚未牢固确立的化学原理时,尤其如此。Cu 可以猝灭基于亚胺的荧光团 DPHP 的荧光,推测是通过络合作用。通过添加 Ru 基 CORM CORM-3,Cu 猝灭的荧光得以恢复。这种方法被报道为一种新的“检测一氧化碳的策略”,提出的机制依赖于 CO 在近生理条件下将 Cu 还原为 Cu(2022 年,94,11298-11306)。该研究仅使用 CORM-3 作为 CO 的来源。据报道,CORM-3 具有非常明显的氧化还原反应性,并且已知除非存在强亲核试剂,否则它不会在水溶液中释放 CO。为了评估 DPHP-Cu(II)混合物对 CORM-3 的荧光响应是否真正通过检测 CO,我们报告了使用纯 CO 和 CORM-3 的实验结果。我们证实了报道的 DPHP-Cu(II)对 CORM-3 的响应,但对纯 CO 气体没有响应。此外,我们没有观察到 DPHP 对 CO 与硫化物物种的报道选择性。沿着这条线,我们还发现还原剂如抗坏血酸能够诱导与 CORM-3 相同的荧光开启。因此,DPHP-Cu(II)系统不是 CO 探针,不能用于研究 CO 生物学。由此得出的结论是,在开发 CO 探针时,未来的工作必须使用不仅仅是具有化学反应性的“CO 供体”作为 CO 源。特别重要的是,要确认“CO 探针”使用纯 CO 气体或另一种 CO 源检测 CO 的能力。