Revalidation of recombinant aequorin as a light emission standard: Estimation of specific activity of Gaussia luciferase
Revalidation of recombinant aequorin as a light emission standard: Estimation of specific activity of Gaussia luciferase
复制标题
DOI：
10.1016/j.bbrc.2018.11.015
复制
发表时间：
2018-12-09
影响因子：
3.1
通讯作者：
Hojo, Hironobu
中科院分区：
文献类型：
--
作者：
Inouye, Satoshi;Hojo, Hironobu
为了验证重组水杉蛋白（reAequorin）作为光发射标准物的有效性，通过氨基酸组成分析确定了高纯度水杉蛋白的蛋白浓度，并通过460 nm处与280 nm处吸光度峰的比值确认了活性水杉蛋白的存在。发光强度与蛋白浓度的高度相关表明，reAequorin可作为荧光素酶发光特性研究的发光标准，并可用于评价光度计的检测灵敏度。具有I-max的高斯荧光素酶的比活性是reAequorin的7.5倍，为3.8 × 10（16）量子/mg蛋白。(C) 2018爱思唯尔公司版权所有。
To validate the use of recombinant aequorin (reAequorin) as a light emission standard, the protein concentrations of highly purified reAequorin were determined by amino acid composition analysis, and the presence of active reAequorin was confirmed by the ratio of absorbance peak at 460 nm to that at 280 nm. The high correlation of the luminescence intensity with the protein concentration showed that reAequorin could be used for a light emission standard to study the luminescence properties of luciferases and to evaluate the detection sensitivity of luminometers. The specific activity of Gaussia luciferase with I-max was 7.5-fold higher than that of reAequorin and was calculated to be 3.8 x 10(16) quanta/mg protein. (C) 2018 Elsevier Inc. All rights reserved.