Synthesis and characterization of two fluorescent sulfhydryl reagents.
Synthesis and characterization of two fluorescent sulfhydryl reagents.
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DOI：
10.1021/bi00745a019
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发表时间：
1973-10
期刊：
影响因子：
2.9
通讯作者：
E. Hudson;G. Weber
中科院分区：
文献类型：
--
作者：
E. Hudson;G. Weber
现在，在蛋白质研究中使用荧光探针已是家喻户晓。自从荧光素异氰酸酯(Creech和Jones，1941)和丹磺酰氯(Weber，1952)被引入以来，各种共价和非共价探针被开发出来并用于阐明蛋白质的各种性质(Steiner和Edelhoch，1962；Stryer，1968；Brand and Gohlke，1972)。然而，除了几个例外，来自伊利诺伊大学厄巴纳化学科学学院生物化学系的f，伊利诺伊州61801。1973年4月23日收到。这些研究得到了来自美国公共卫生服务的国家普通医学科学研究所的Grant GM 11223的支持。目前地址：威斯康星州沃基沙卡罗尔学院化学系。53186。产生光稳定性共价衍生物的蛋白质中的干基。本文介绍了模型化合物在不同溶剂中的合成、荧光光谱、量子产率和寿命。下面的一篇论文将报道蛋白质结合物的一些一般性质。共价探针是非特异性的，它们与几种不同的氨基酸侧链反应。这导致了异质标记和随后的荧光特性测量，这些测量来自许多不同的位置，因此是平均的。这种特异性的缺乏明显地限制了用这种探针进行测量的应用和解释。我们希望报告几种新的荧光试剂的合成和表征，这些试剂结合了碘乙酰胺对硫基的反应活性和萘磺酸的光谱性质。该试剂合成简单，易溶于水，以晶体形式长期稳定，并与蛋白质形成稳定的偶联物。像其他的萘胺磺酸一样，它们
TA he use of fluorescent probes in protein studies is now well established. Since the introduction of fluorescein isocyanate (Creech and Jones, 1941) and dansyl chloride (Weber, 1952), a variety of covalent and noncovalent probes have been developed and applied to the elucidation of various properties of proteins (Steiner and Edelhoch, 1962; Stryer, 1968; Brand and Gohlke, 1972). With a few exceptions, however, the f From the Biochemistry Department, School of Chemical Sciences, University of Illinois, Urbana, Illinois 61801. Received April 23, 1973. These studies were supported by Grant GM 11223 from the National Institute of General Medical Sciences, U. S. Public Health Service. t Present address: Chemistry Department, Carroll College, Waukesha, Wis. 53186. dryl groups in proteins yielding photostable covalent deriva-tives. The synthesis, fluorescence spectra, quantum yields, and lifetimes of model compounds in various solvents are pre-sented in this paper. A following paper will report some of the general properties of the protein conjugates. covalent probes have been nonspecific;/.<?., they react with several different amino acid side chains. This results in hetero-geneous labeling and thesubsequent measurements of fluorescent properties arise from, and are therefore averaged over, a number of different sites. This lack of specificity clearly limits the application and interpretation of the measurements with such probes.We wish to report in this paper the synthesis and character-ization of several new fluorescent reagents which combine the reactivity of iodoacetamide toward sulfhydryl groups with the spectral properties of the naphthalenesulfonic acids. The re-agents are easily synthesized, water soluble, stable for long periods of time in crystallineform, and form stable conjugates with proteins. Like other naphthylaminesulfonic acids, they