一种简单的慢速给药-快速淬灭装置的构建与测试。
The construction and testing of a simple, slow delivery-rapid quench apparatus.
作者信息
Ray W J, Long J W
出版信息
Biochemistry. 1976 Sep 7;15(18):3990-3. doi: 10.1021/bi00663a013.
A simple, inexpensive, slow delivery-rapid quench apparatus is described. The apparatus can be used to mix small volumes (about 50 mul) of equilibrium mixtures of enzyme-substrate and enzyme-product complexes with a quenching solution, ideally to inactivate the enzyme more rapidly than such complexes can be inconverted. The efficiency of the apparatus is tested by (a) injecting basic solutions of an indicator dye into acid and observing the length of the unbleached plume of dye produced at the delivery tip and (b) by forming an enzyme-substrate complex with 32P-labeled phosphoglucomutase and measuring the extent of label transfer prior to inactivation by the quenching solution. Problems that may be encountered in attempts to inactivate equilibrium mixtures of enzyme-substrate and enzyme-product complexes without producing quenching artifacts are considered.
描述了一种简单、廉价的慢速输送-快速淬灭装置。该装置可用于将少量（约50微升）酶-底物和酶-产物复合物的平衡混合物与淬灭溶液混合，理想情况下，其灭活酶的速度要比这些复合物转化的速度更快。通过以下方式测试该装置的效率：(a) 将指示剂染料的碱性溶液注入酸中，并观察在输送尖端产生的未漂白染料羽流的长度；(b) 用32P标记的磷酸葡萄糖变位酶形成酶-底物复合物，并在被淬灭溶液灭活之前测量标记转移的程度。还考虑了在试图灭活酶-底物和酶-产物复合物的平衡混合物而不产生淬灭假象时可能遇到的问题。
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