N,S-Bis-Fmoc-glutathione(DiFMOC-G) which has been synthesized and characterized is a very potent competitive inhibitor of mammalian glyoxalase II activity(K, =0.75 M, calfliver glyoxalase II). Among other enzymes involved in glutathione metabolism, such as glutathione S-transferase, glutathione reductase, and glutathione peroxidase, only glutathione S-transferase is inhibited to a small extent by N,S-Bis-Fmoc-glutathione [2]. The thiocarbonate derivatives of glutathione, which bind very strongly to glyoxalase II and serve as effective competitive inhibitors of the enzyme, also serve as weak substrates for glyoxalase II[1].For example, the KM values for p-NCB-G and for N,S-Bis-Fmoc-glutathione are 4 µM and 1.5 µM, respectively, when tested with pure calf-liver glyoxalase II.The relative V,max.values, compared with the V,max.with SLG as substrate, are 0.125% and 0.45%, for pNCB-G and N,S-Bis-Fmoc-glutathione, respectively[1].
Embryonic neural cells undergo rather major elaborations of network connections within a few days of culturing in vitro. These networks, resulting from the extension of processes elaborated by the cells, require extensive synthesis ofamicrotubularcytoskeleton [3]. These neurons show profound effects from the N,S-Bis-Fmoc-glutathione diester treatment; extending neurites have contracted or disappeared and the cells, in some cases, have begun to disintegrate. After 47-72 h exposure to the N,S-Bis-Fmoc-glutathione diester, all identifiable cells have become globular in shape[2].
References:
[1]: Norton SJ, Elia AC, et,al. Inhibitors and inhibition studies of mammalian glyoxalase II activity. Biochem Soc Trans. 1993 May;21(2):545-9. doi: 10.1042/bst0210545. PMID: 8359529.
[2]:EliaAC,ChyanMK,et,al.N,S-bis-fluorenylmethoxycarbonylglutathione: a new, very potent inhibitor of mammalian glyoxalase II. Biochem Mol Biol Int. 1995 Apr;35(4):763-71. PMID: 7627127.
[3]: Alberts, B., Bray, D. et,al. (2014). Essential Cell Biolog (4th ed.). W.W. Norton & Company. https://doi.org/10.1201/9781315815015
N,S-Bis-Fmoc-glutathione(DiFMOC-G) 已被合成和表征，是哺乳动物乙二醛酶 II 活性（K，=0.75 M，小牛肝乙二醛酶 II）的一种非常有效的竞争性抑制剂。在参与谷胱甘肽代谢的其他酶中，如谷胱甘肽 S-转移酶、谷胱甘肽还原酶和谷胱甘肽过氧化物酶，只有谷胱甘肽 S-转移酶受到 N,S-Bis-Fmoc-谷胱甘肽的小程度抑制[2]< /sup>。谷胱甘肽的硫代碳酸酯衍生物与乙二醛酶 II 结合非常牢固，是该酶的有效竞争性抑制剂，也是乙二醛酶 II[1] 的弱底物。例如，p 的 KM 值-NCB-G 和 N,S-Bis-Fmoc-谷胱甘肽分别为 4 μM 和 1.5 μM，当用纯小牛肝乙二醛酶 II 测试时。相对 V,max.values，与 V,max.with 相比SLG作为底物，pNCB-G和N,S-Bis-Fmoc-glutathione分别为0.125%和0.45%[1]。
胚胎神经细胞在体外培养的几天内经历了相当重要的网络连接精细化。这些网络是由细胞精心设计的过程的扩展产生的，需要大量合成微管细胞骨架[3]。这些神经元显示出 N,S-双-Fmoc-谷胱甘肽二酯治疗的深远影响；延伸的神经突收缩或消失，在某些情况下，细胞开始分解。暴露于 N,S-Bis-Fmoc-谷胱甘肽二酯 47-72 小时后，所有可识别的细胞都变成了球形[2]。