停流X射线散射：天冬氨酸转氨甲酰酶的解离
Stopped-flow x-ray scattering: the dissociation of aspartate transcarbamylase.
作者信息
Moody M F, Vachette P, Foote A M, Tardieu A, Koch M H, Bordas J
出版信息
Proc Natl Acad Sci U S A. 1980 Jul;77(7):4040-3. doi: 10.1073/pnas.77.7.4040.
Abstract
A combination of stopped-flow and x-ray scattering techniques was used to study the dissociation of aspartate transcarbamylase (carbamoylphosphate:L-aspartate carbamoyltransferase, EC 2.1.3.2) with a 2:1 excess of p-chloromercuribenzenesulfonic acid (the ratio being calculated on a basis of reactive sites), in the presence and absence of the transition state analogue N-(phosphonacetyl)-L-aspartate. At 10 mg of protein per ml, the scattering curves allowed some details of the reaction to be followed with a time resolution down to 1 sec. The curves showed not only the dissociation of the enzyme complex but also the formation of the subunits. These results show that, with present facilities, x-ray scattering could be used to study dissociation or reassociation reactions with a time resolution of the order of 100 msec.
摘要
采用停流技术与X射线散射技术相结合的方法，在有和没有过渡态类似物N-（膦酰基乙酰基）-L-天冬氨酸存在的情况下，研究了天冬氨酸转氨甲酰酶（氨甲酰磷酸：L-天冬氨酸氨甲酰转移酶，EC 2.1.3.2）与2:1过量的对氯汞苯磺酸（该比例基于反应位点计算）的解离。在每毫升10毫克蛋白质的情况下，散射曲线使得能够以低至1秒的时间分辨率追踪反应的一些细节。这些曲线不仅显示了酶复合物的解离，还显示了亚基的形成。这些结果表明，利用现有设备，X射线散射可用于研究时间分辨率约为100毫秒的解离或重新缔合反应。
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