DECAMERS OBSERVED IN CRYSTAL OF BOVINE PANCREATIC TRYPSIN INHIBITOR

牛胰蛋白酶抑制剂晶体中观察到的十聚体

• 批准号: 6205777
• 负责人: ALEX WLODAWER
• 金额: --
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-09-01 至 2000-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6205777
• 关键词: biological products; biomedical resource; musculoskeletal system; proteins; spectrometry; synchrotrons

Profilin is a ubiquitous 15kD protein required for the normal distribution of filamentous actin structures in vivo. Profilin binds PIP2, suggesting a possible link to signal transduction pathways. Profilins from different organisms have limited sequence homology, but highly homologous 3D structures, indicating an evolutionary pressure to maintain the protein fold. It has been the goal of our laboratory to combine crystallography and in vivo studies to examine the role of profilin in the cell. Two profilin data sets were collected at X9B. Crystals of P2 profilin, (a profilin isoform of Acanthamoeba castellani), which diffracted only to 4.0A using a rotating anode and diffracted to 2.0A at X9B, has a space group of F432 and unit cell parameters of a=b=c=241.28, a=b=g=90o. The atomic structure of P2 profilin will aid in the creation of mutants with which to study the specific lipid binding activity of this protein. Crystals of Saccharomyces cerevisiae profilin which diffracted weakly using a rotating anode, diffracted to 3.0 A at X9B. The S. Cerevisiae crystals are of the P432 space group with unit cell parameters of a=b=c=127.66, a=b=g=90o and were successfully reduced and scaled (Rmerge of 5.8%). We have now prepared a new crystal form of yeast profilin (P65; a=b=58.1, c=151.3E) and have refined the structure to 2.3E resolution. This structure is being used to design specific mutants that will probe the in vivo role of actin and proline rich-ligand binding. Furthermore, we have recently solved the structure of human profilin bound to a decamer of L-proline which gives the first structural information about this interaction that is responsible for the intra-cellular localization of profilin.

Profilin是一种普遍存在的15 kD蛋白质，是正常人 丝状肌动蛋白结构在体内的分布。 Profilin结合 PIP 2，提示可能与信号转导途径有关。 来自不同生物体的Profilins具有有限的序列同源性，但 高度同源的三维结构，表明进化压力 来维持蛋白质折叠。 这是我们实验室的目标 结合联合收割机和体内研究， profilin in in the cell细胞. 在X9 B收集两个profilin数据集。 P2 profilin的晶体（阿米巴原虫的profilin同种型 castellani），其使用旋转阳极仅衍射至4.0A， 在X9 B处衍射至2.0A，具有F432空间群和晶胞 参数a=B=c=241.28，a=B=g= 90 °。 P2的原子结构 profilin将有助于创造突变体， 这种蛋白质的特异性脂质结合活性。 晶体 酿酒酵母profilin的衍射弱，使用 旋转阳极，在X9 B处衍射至3.0 A。 色葡萄酿酒酵母 晶体属于P432空间群，晶胞参数为 a=B=c=127.66，a=B=g= 90 o，并成功地进行了缩小和缩放 （5.8%）。 我们现在准备了一种新的酵母晶体 profilin（P65; a=B=58.1，c=151.3E），并已将结构细化为 2.3E分辨率。 这种结构被用来设计特定的 将探测肌动蛋白和脯氨酸在体内作用的突变体 富配体结合。 此外，我们最近还解决了 与L-脯氨酸十聚体结合的人profilin的结构， 给出了关于这种相互作用的第一个结构信息， 负责profilin的细胞内定位。