Guo Hao, Ding Li, Zhang Tian-Jie, Mao Yan-Li
School of Physics and Electronics, Henan University, Kaifeng 475004, China.
Guang Pu Xue Yu Guang Pu Fen Xi. 2013 May;33(5):1215-9.
Dithiobis-succinimidyl propionate (DTSP), an important homobifunctional crosslinker, has been widely used for the covalent immobilization of proteins onto solid supports by amine coupling. In the present study, adsorption of DTSP on vacuum-deposited gold island films was analyzed by means of surface-enhanced infrared absorption spectroscopy (SEIRAS). For the sake of a reliable assignment of the vibrational spectra, IR intensity of the adsorption model of TSP on one gold surface was calculated using density functional theory (DFT) at the Beck' s three-parameter Lee-Yang-Parr (B3LYP) level with the LANL2DZ basis set. SEIRAS and multiple-angle-of-incidence polarization infrared reflection-absorption spectroscopy indicated that TSP is arranged orderly in a tilted fashion with a dihedral angle of 65 degrees between the plane of succinimidyl ring and the gold surface. The binding kinetics revealed that that the time constant of self-assembly of the TSP layer is 220 sec. Furthermore, the coupling process of amino-nitrilotriacetic acid (ANTA) with surface-bound TSP monolayer was monitored in situ by SEIRAS. Three negative bands observed at 1 807, 1 776, and 1 728 cm(-1) respectively provided direct evidence for the reaction of the succinimidyl ester. The appearance of one intense band at 1 566 cm(-1) gave a clear support for the presence of the cross-link between ANTA and TSP. We hope that the results in current investigation will contribute to the better understanding of properties of DTSP and related reactions at the molecular level.
二硫代双琥珀酰亚胺丙酸酯(DTSP)是一种重要的同双功能交联剂,已被广泛用于通过胺偶联将蛋白质共价固定在固体支持物上。在本研究中,通过表面增强红外吸收光谱(SEIRAS)分析了DTSP在真空沉积金岛膜上的吸附情况。为了可靠地归属振动光谱,使用密度泛函理论(DFT)在Beck三参数Lee-Yang-Parr(B3LYP)水平和LANL2DZ基组下计算了TSP在一个金表面上的吸附模型的红外强度。SEIRAS和多角度入射偏振红外反射吸收光谱表明,TSP以倾斜方式有序排列,琥珀酰亚胺环平面与金表面之间的二面角为65度。结合动力学表明,TSP层自组装的时间常数为220秒。此外,通过SEIRAS原位监测了氨基三乙酸(ANTA)与表面结合的TSP单层的偶联过程。分别在1807、1776和1728cm-1处观察到的三个负带为琥珀酰亚胺酯的反应提供了直接证据。在1566cm-1处出现的一个强带清楚地支持了ANTA与TSP之间交联的存在。我们希望当前研究的结果将有助于在分子水平上更好地理解DTSP的性质和相关反应。