Zhang Yanjie, Trabbic-Carlson Kimberly, Albertorio Fernando, Chilkoti Ashutosh, Cremer Paul S
Department of Chemistry, Texas A&M University, 3255 TAMU, College Station, Texas 77843, USA.
Biomacromolecules. 2006 Jul;7(7):2192-9. doi: 10.1021/bm060254y.
The kinetics of aqueous two-phase system (ATPS) formation for elastin-like polypeptides (ELP) with defined chemical composition and chain length was investigated by dark field microscopy in an on-chip format with a linear temperature gradient. Scattering intensities from peptide solutions in the presence and absence of sodium dodecyl sulfate (SDS) were recorded as a function of temperature and time, simultaneously. It was found that the formation of the ATPS for three ELPs of different molecular weights (36 075, 59 422, and 129 856 Da) in the absence of SDS followed a coalescence mechanism, and the rate constant and activation energy were independent of chain length. With the introduction of SDS into the ELP solutions, the rate constants were attenuated more strongly with increasing chain length. Moreover, the coalescence process in the presence of SDS showed non-Arrhenius kinetics as a function of temperature. For the two shorter ELPs, ATPS formation occurred via coalescence at all SDS concentrations and temperatures investigated. On the other hand, the coalescence process was greatly suppressed for the longest ELP at elevated temperatures and higher SDS concentrations. Under these circumstances, ATPS formation was forced to proceed via a mixed Ostwald ripening and coalescence mechanism.
通过暗场显微镜在具有线性温度梯度的芯片上对具有确定化学组成和链长的类弹性蛋白多肽(ELP)形成水相两相系统(ATPS)的动力学进行了研究。同时记录了在有和没有十二烷基硫酸钠(SDS)的情况下肽溶液的散射强度随温度和时间的变化。发现在没有SDS的情况下,三种不同分子量(36075、59422和129856 Da)的ELP形成ATPS遵循聚并机制,速率常数和活化能与链长无关。随着SDS引入ELP溶液中,速率常数随着链长增加而衰减得更强。此外,在SDS存在下的聚并过程表现出非阿累尼乌斯动力学随温度的变化。对于两个较短的ELP,在所研究的所有SDS浓度和温度下,ATPS形成均通过聚并发生。另一方面,对于最长的ELP,在高温和较高SDS浓度下,聚并过程受到极大抑制。在这些情况下,ATPS形成被迫通过混合奥斯特瓦尔德熟化和聚并机制进行。