CAJ | 학술논문

采用全原子分子动力学方法模拟了反相色谱分离蛋白质的吸附和洗脱过程.采用表面键合C4烷基链的硅胶基质作为反相色谱介质和细胞色素C作为蛋白质模型,模拟蛋白质在反相色谱分离过程中的构象变化.结果显示:在吸附过程中,蛋白质在释放出表面结合水分子的同时置换出色谱介质表面的水分子.与其在溶液中的天然构象相比,吸附态蛋白质的构象发生改变.在洗脱过程中,随着溶剂从水切换到甲醇,甲醇取代水分子包覆在介质和蛋白质表面,将蛋白质从介质表面置换下来.分子模拟的结果再现了有关反相色谱"优先水化"的吸附机制,并从分子水平上展现了吸附和洗脱过程中蛋白质、色谱介质和溶剂之间相互作用,对反相色谱介质设计和过程优化提供了参考.
채용전원자분자동역학방법모의료반상색보분리단백질적흡부화세탈과정.채용표면건합C4완기련적규효기질작위반상색보개질화세포색소C작위단백질모형,모의단백질재반상색보분리과정중적구상변화.결과현시:재흡부과정중,단백질재석방출표면결합수분자적동시치환출색보개질표면적수분자.여기재용액중적천연구상상비,흡부태단백질적구상발생개변.재세탈과정중,수착용제종수절환도갑순,갑순취대수분자포복재개질화단백질표면,장단백질종개질표면치환하래.분자모의적결과재현료유관반상색보"우선수화"적흡부궤제,병종분자수평상전현료흡부화세탈과정중단백질、색보개질화용제지간상호작용,대반상색보개질설계화과정우화제공료삼고.
This study presents a molecular dynamics simulation of protein separation by reverse phase liquid chromatography(RPLC)at all-atom level,using silica gel surface with C4 compounds and cytochrome C as representative adsorbent and protein,respectively.The conformational transition of cytochrome C was monitored during the aqueous adsorption onto the adsorbent and elution with methanol.It was shown that during the adsorption,in which protein was attracted to the adsorbent surface,a simultaneous exclusion of water molecules occurred both at the adsorbent surface and the protein.A conformational transition of the protein conformation that led to the exposure of the hydrophobic amino acid residues,as compared to its native conformation in aqueous phase,was also displayed.During the elution by methanol,the replacement of water molecule by methanol occurred on both the surface of adsorbent and the adsorbed protein.As a result,the adsorbed protein was replaced by methanol.Above-mentioned simulation results reproduced the preferential hydration mechanism of the adsorption by RPLC and provided a molecular insight into the interaction among protein,adsorbent and the solvents at all-atom level,which is helpful for the design of new adsorbent and the optimization of RPLC process.