Design of Coelenterazine Analogue to Reveal Bioluminescent Reaction of Human Serum Albumin
Design of Coelenterazine Analogue to Reveal Bioluminescent Reaction of Human Serum Albumin
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DOI：
10.5772/intechopen.97232
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发表时间：
2021-04
期刊：
影响因子：
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通讯作者：
Ryohei Nishihara;K. Niwa;Tatsunosuke Tomita;R. Kurita
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作者：
Ryohei Nishihara;K. Niwa;Tatsunosuke Tomita;R. Kurita
本章介绍了我们设计的一种咪唑并吡嗪酮类荧光素分子HuLumino 1，并对其发光性质进行了研究。这种荧光素被设计成通过人血清白蛋白（HSA）而不是来自发光生物体的荧光素酶产生生物发光。HuLumino 1是通过将甲氧基封端的烷基链修饰到C-6位并消除腔肠素C-8位的苄基而开发的。为了阐明HSA发光的基础，使用校准的光度计研究了HuLumino 1/HSA对的详细动力学性质。仅在HSA存在下观察到HuLumino 1的酶促氧化。使用HuLumino 1的HSA定量实验的结果与酶联免疫吸附测定的结果的差异小于5%，这表明HuLumino 1可用于血清样品中HSA水平的定量分析而无需任何预处理。这些结果证明了腔肠素类似物作为生物发光试剂检测非标记蛋白质的优势，其通常不起酶的作用。
This chapter describes the design of an imidazopyrazinone-type luciferin named as HuLumino1 by us and investigation of its luminescence properties. This luciferin was designed to generate bioluminescence by human serum albumin (HSA) rather than by luciferase derived from luminous organisms. HuLumino1 was developed by modifying a methoxy-terminated alkyl chain to the C-6 position and eliminating a benzyl group at the C-8 position of coelenterazine. To clarify the basis of light emission by HSA, the detailed kinetic properties of the HuLumino1/HSA pair were investigated using a calibrated luminometer. The enzymatic oxidation of HuLumino1 was observed only in the presence of HSA. Results of HSA quantification experiments using HuLumino1 agreed with less than 5% differences with those of enzyme-linked immunosorbent assays, suggesting HuLumino1 could be used for quantitative analysis of HSA levels in serum samples without any pretreatments. These results demonstrate the advantages of the coelenterazine analogue as a bioluminescence reagent to detect non-labeled proteins, which generally do not function as enzymes.