MACCHESS CONSORTIUM FOR LARGE MACROMOLECULAR STRUCTURES: HERPES VIRUS

MACCHESS 大分子结构联盟：疱疹病毒

• 批准号: 6491123
• 负责人: JAMES M HOGLE
• 金额: $ 14.27万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-08-15 至 2002-08-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6491123
• 关键词: bioimaging /biomedical imaging; biological products; biomedical resource; proteins; radiography; structural biology

Penicillopepsin (EC 3.2.23.X) is an aspartic proteinase isolated from the fungus Penicillium janthinellum. In preparation for the data collection at CHESS, crystals were grown of the native enzyme and of its complexes with two cyclic phosphonate inhibitors, methyl[cyclo-7[(2R)-((N-valyl) amino)-2-(hydroxyl-(1S)-1-methyoxycarbonyl-2-phenyl-ethoxy)phosphinyloxy (PP6) and methyl cyclo[(2S)-2-[[(1R)-1-(N-(L-N-(3-methylbutanoyl)valyl-L-aspartyl)amino)- (PP7). The phosphonate inhibitors act as mimics of the transition state in the cleavage of good substrates. Cyclization of the inhibitor reduces its flexibility and lowers the entropic cost of binding to the enzyme. At CHESS, crystals were transferred to solutions of the mother liquor containing increasing amounts of the cryoprotectant, glycerol. The final solution contained 20% glycerol, 40% saturated ammonium sulfate and 100 mM sodium acetate at pH 4.6. Data from five crystals, one of the native and two of each complex, were collected at 100 K with X-rays of wavelength 0.919
on the ADSC Quantum-4 CCD detector at station F1. The native data were processed to 0.95
: 415771 data were measured for 143710 unique reflections (Rmerge = 6.8%), 88.1% complete. The data from one crystal of each complex were processed to 0.90
resolution. For PP6, 614135 measurements were made of 174924 unique reflections (Rmerge = 4.2%), 89.7% complete; for PP7, 459707 measurements were made of 156276 unique reflections (Rmerge = 5.3%), 80.1% complete to 0.90
. The three models are currently being refined against these data. The crystallographic residuals are 11.2% for the native, 11.8% for the PP6 complex and 11.0% for the PP7 complex.

青霉蛋白酶 (EC 3.2.23.X) 是一种分离的天冬氨酸蛋白酶 来自真菌Penicillium janthinellum。 正在准备数据 在 CHESS 收集的晶体中生长出天然酶和 它与两种环状膦酸酯抑制剂的复合物， 甲基[环-7[(2R)-((N-缬氨酰基) 氨基)-2-(羟基-(1S)-1-甲氧基羰基-2-苯基-乙氧基)氧膦氧基 (PP6) 和甲基 环[(2S)-2-[[(1R)-1-(N-(L-N-(3-甲基丁酰基)缬氨酰基-L-天冬氨酰基)氨基)- （PP7）。 膦酸盐抑制剂充当转变的模拟物 良好底物裂解状态。 环化 抑制剂降低了其灵活性并降低了熵成本 与酶结合。 在 CHESS 中，晶体被转移到 母液溶液中含有越来越多的 冷冻保护剂、甘油。 最终溶液含有20%甘油， 40% 饱和硫酸铵和 100 mM 乙酸钠，pH 4.6。 来自五种晶体的数据，其中一种是天然晶体，另一种是复合晶体， 在 ADSC 上使用波长 0.919 的 X 射线在 100 K 下收集 F1 站的 Quantum-4 CCD 探测器。 原生数据已处理 至 0.95：针对 143710 个独特反射测量了 415771 个数据 （重新合并 = 6.8%），完成 88.1%。 每个晶体的数据 复合物被处理至0.90分辨率。 对于 PP6，614135 测量由 174924 个独特的反射（Rmerge = 4.2%）组成， 完成89.7%；对于 PP7，156276 进行了 459707 次测量 独特的反射（Rmerge = 5.3%），80.1% 完成到 0.90 。这 目前正在根据这些数据完善三个模型。 这 天然晶体残留为 11.2%，PP6 为 11.8% 复合物，PP7 复合物为 11.0%。