Gruys K J, Schuster S M
J Inorg Biochem. 1986 Jun;27(2):75-84. doi: 10.1016/0162-0134(86)80009-5.
The two chelation isomers of CrITP, gamma-monodentate and beta, gamma-bidentate CrITP, as well as the diastereomers of beta, gamma-bidentate CrITP were synthesized, isolated, and characterized. Synthesis of these complexes was done using pH titration methods similar to that described by Cleland [W.W. Cleland, Methods Enzymol. 87, 159 (1982)], and separation of the two chelation isomers was accomplished with DEAE-sephadex A-25 using 0-0.3 N linear HCl gradient. Diastereomer separation (analytical and preparative scales) of beta, gamma-bidentate CrITP using reverse-phase high-performance liquid chromatography, and then analysis of the diastereomers with circular dichroism spectroscopy, shows four diastereomers that exist as two pairs of mirror-image isomers, similar to the four diastereomers of beta, gamma-bidentate CrATP as presented by Dunaway-Mariano and Cleland [D. Dunaway-Mariano and W.W. Cleland, Biochemistry 19, 1496 (1980)]. Reverse-phase high-performance liquid chromatography analysis of gamma-monodentate CrITP shows the presence of two major peaks, both of which convert to beta, gamma-bidentate CrITP upon incubation at pH 6.0 for 1 hr.
合成、分离并表征了CrITP的两种螯合异构体，即γ-单齿和β,γ-双齿CrITP，以及β,γ-双齿CrITP的非对映异构体。这些配合物的合成采用了类似于Cleland [W.W. Cleland, Methods Enzymol. 87, 159 (1982)] 所述的pH滴定法，两种螯合异构体的分离是使用DEAE-葡聚糖A-25，通过0 - 0.3 N线性HCl梯度实现的。使用反相高效液相色谱对β,γ-双齿CrITP进行非对映异构体分离（分析规模和制备规模），然后用圆二色光谱分析非对映异构体，结果显示存在四个非对映异构体，它们以两对镜像异构体的形式存在，类似于Dunaway-Mariano和Cleland [D. Dunaway-Mariano和W.W. Cleland, Biochemistry 19, 1496 (1980)] 报道的β,γ-双齿CrATP的四个非对映异构体。γ-单齿CrITP的反相高效液相色谱分析显示有两个主要峰，在pH 6.0孵育1小时后，这两个峰均转化为β,γ-双齿CrITP。
