McNaughton Lynn, Li Zhong, Van Roey Patrick, Hanes Steven D, LeMaster David M
Wadsworth Center, New York State Department of Health, School of Public Health, University at Albany, Empire State Plaza, Albany, NY 12201, USA.
Biochim Biophys Acta. 2010 Jul;1804(7):1537-41. doi: 10.1016/j.bbapap.2010.03.005. Epub 2010 Mar 18.
Ess1 is a peptidyl prolyl cis/trans isomerase that is required for virulence of the pathogenic fungi Candida albicans and Cryptococcus neoformans. The enzyme isomerizes the phospho-Ser-Pro linkages in the C-terminal domain of RNA polymerase II. Its human homolog, Pin1, has been implicated in a wide range of human diseases, including cancer and Alzheimer's disease. Crystallographic and NMR studies have demonstrated that the sequence linking the catalytic isomerase domain and the substrate binding WW domain of Pin1 is unstructured and that the two domains are only loosely associated in the absence of the substrate. In contrast, the crystal structure of C. albicans Ess1 revealed a highly ordered linker that contains a three turn alpha-helix and extensive association between the two tightly juxtaposed domains. In part to address the concern that the marked differences in the domain interactions for the human and fungal structures might reflect crystal lattice effects, NMR chemical shift analysis and 15N relaxation measurements have been employed to confirm that the linker of the fungal protein is highly ordered in solution. With the exception of two loops within the active site of the isomerase domain, the local backbone geometry observed in the crystal structure appears to be well preserved throughout the protein chain. The marked differences in interdomain interactions and linker flexibility between the human and fungal enzymes provide a structural basis for therapeutic targeting of the fungal enzymes.
Ess1是一种肽基脯氨酰顺反异构酶,是致病性真菌白色念珠菌和新型隐球菌致病力所必需的。该酶使RNA聚合酶II C末端结构域中的磷酸丝氨酸-脯氨酸连接发生异构化。其人类同源物Pin1与多种人类疾病有关,包括癌症和阿尔茨海默病。晶体学和核磁共振研究表明,连接Pin1催化异构酶结构域和底物结合WW结构域的序列是无结构的,并且在没有底物的情况下,这两个结构域仅松散结合。相比之下,白色念珠菌Ess1的晶体结构显示出一个高度有序的连接子,其中包含一个三圈α螺旋,并且两个紧密相邻的结构域之间存在广泛的关联。为了部分解决关于人类和真菌结构在结构域相互作用上的显著差异可能反映晶格效应的担忧,已采用核磁共振化学位移分析和15N弛豫测量来证实真菌蛋白的连接子在溶液中是高度有序的。除了异构酶结构域活性位点内的两个环之外,晶体结构中观察到的局部主链几何形状在整个蛋白质链中似乎都得到了很好的保留。人类和真菌酶在结构域间相互作用和连接子灵活性上的显著差异为真菌酶的治疗靶向提供了结构基础。