Toyama Prefectural Institute for Pharmaceutical Research, Imizu 939-0363, Toyama, Japan.
Department of Medical Chemistry, Kansai Medical University, Hirakata 573-1010, Osaka, Japan.
Molecules. 2022 Jul 30;27(15):4889. doi: 10.3390/molecules27154889.
Molecular interaction analysis is an essential technique for the study of biomolecular functions and the development of new drugs. Most current methods generally require manipulation to immobilize or label molecules, and require advance identification of at least one of the two molecules in the reaction. In this study, we succeeded in detecting the interaction of low-molecular-weight (LMW) compounds with a membrane protein mixture derived from cultured cells expressing target membrane proteins by using the size exclusion chromatography-mass spectrometry (SEC-MS) method under the condition of 0.001% lauryl maltose neopentyl glycol as detergent and atmospheric pressure chemical ionization. This method allowed us to analyze the interaction of a mixture of medicinal herbal ingredients with a mixture of membrane proteins to identify the two interacting ingredients. As it does not require specialized equipment (e.g., a two-dimensional liquid chromatography system), this SEC-MS method enables the analysis of interactions between LMW compounds and relatively high-expressed membrane proteins without immobilization or derivatization of the molecules.
分子相互作用分析是研究生物分子功能和开发新药的重要技术。目前大多数方法通常需要进行操作以固定或标记分子，并至少需要预先鉴定反应中的两种分子之一。在这项研究中，我们成功地使用尺寸排阻色谱-质谱（SEC-MS）法，在 0.001%月桂基麦芽糖新戊二醇作为洗涤剂和大气压化学电离的条件下，检测到低分子量（LMW）化合物与来源于表达目标膜蛋白的培养细胞的膜蛋白混合物之间的相互作用。该方法允许我们分析药用草药成分混合物与膜蛋白混合物之间的相互作用，以鉴定两种相互作用的成分。由于它不需要特殊的设备（例如二维液相色谱系统），因此这种 SEC-MS 方法可以在不固定或衍生分子的情况下分析 LMW 化合物与相对高表达的膜蛋白之间的相互作用。
