Pomes A, Kempner E, Rohrer S
Department of Cell and Biochemical Physiology, Merck Research Laboratories, Rahway, NJ 07065, USA.
Biochim Biophys Acta. 1997 May 23;1339(2):233-8. doi: 10.1016/s0167-4838(97)00006-x.
A high-affinity avermectin binding site from Drosophila melanogaster head membranes was subjected to target size analysis by radiation inactivation in order to determine the functional unit size. Using the [3H]ivermectin binding assay to assess ligand binding activity, the target size was determined to be 44.3 +/- 3.9 kDa. This result suggests that the size of the functional unit required for high-affinity ligand binding is a monomer. The membrane-associated acetylcholinesterase present in the Drosophila head membranes was also analyzed by radiation inactivation and served as a control. By monitoring enzymatic activity, the functional unit size of the Drosophila acetylcholinesterase was determined to be 70 +/- 9.7 kDa. This corresponds to the molecular weight of a dimer composed of a 55 kDa subunit and a 16-18 kDa subunit.
为了确定功能单位大小,对来自黑腹果蝇头部膜的高亲和力阿维菌素结合位点进行了辐射失活的靶标大小分析。使用[3H]伊维菌素结合试验评估配体结合活性,确定靶标大小为44.3±3.9 kDa。该结果表明高亲和力配体结合所需功能单位的大小是一个单体。还通过辐射失活分析了黑腹果蝇头部膜中存在的膜相关乙酰胆碱酯酶,并将其作为对照。通过监测酶活性,确定黑腹果蝇乙酰胆碱酯酶的功能单位大小为70±9.7 kDa。这与由一个55 kDa亚基和一个16 - 18 kDa亚基组成的二聚体的分子量相对应。