Functional Profiling of Recombinant NS3 Proteases from All Four Serotypes of Dengue Virus Using Tetrapeptide and Octapeptide Substrate Libraries*[boxs]
Functional Profiling of Recombinant NS3 Proteases from All Four Serotypes of Dengue Virus Using Tetrapeptide and Octapeptide Substrate Libraries*[boxs]
复制标题
使用四肽和八肽底物文库对登革热病毒所有四种血清型的重组 NS3 蛋白酶进行功能分析*[框]
DOI：
--
复制
发表时间：
2005
影响因子：
4.8
通讯作者：
S. Vasudevan
中科院分区：
文献类型：
--
作者：
Jun Li;S. Lim;David Beer;Viral Patel;D. Wen;Christine Tumanut;D. Tully;Jennifer A. Williams;Jan Jiricek;J. Priestle;Jennifer L. Harris;S. Vasudevan
登革热病毒的双组分 NS2B/NS3 蛋白酶调节的蛋白水解对于病毒复制和感染性病毒颗粒的成熟至关重要。通过使用位置扫描格式的四肽和八肽文库（每个包含 130,321 个底物）表征其底物特异性的差异，解决了来自四种遗传和抗原不同血清型的 NS2B/NS3 蛋白酶之间的功能相似性。基于底物切割偏好的相似性，来自不同血清型的蛋白酶被证明在功能上是同源的。观察到 P1 位点对碱性氨基酸残基 (Arg/Lys) 的强烈偏好，而对 P2-4 位点的偏好顺序为：P2 为 Arg > Thr > Gln/Asn/Lys，P3 为 Lys > Arg > Asn，P4 为 Nle > Leu > Lys > Xaa。主要位点底物特异性针对 P1' 和 P3' 中的小极性氨基酸。相反，P2'和P4'底物位置显示出最小的活性。通过具有最佳和次优底物序列的单底物动力学，确定了 P2 和 P3 氨基酸对基态结合和 P4 过渡态稳定位置的影响。观察到的登革热 NS2B/NS3 特异性与登革热多蛋白中的生理切割位点具有共同特征；然而，所有位点都揭示了以前未被识别的次优序列。
Regulated proteolysis by the two-component NS2B/NS3 protease of dengue virus is essential for virus replication and the maturation of infectious virions. The functional similarity between the NS2B/NS3 proteases from the four genetically and antigenically distinct serotypes was addressed by characterizing the differences in their substrate specificity using tetrapeptide and octapeptide libraries in a positional scanning format, each containing 130,321 substrates. The proteases from different serotypes were shown to be functionally homologous based on the similarity of their substrate cleavage preferences. A strong preference for basic amino acid residues (Arg/Lys) at the P1 positions was observed, whereas the preferences for the P2-4 sites were in the order of Arg > Thr > Gln/Asn/Lys for P2, Lys > Arg > Asn for P3, and Nle > Leu > Lys > Xaa for P4. The prime site substrate specificity was for small and polar amino acids in P1′ and P3′. In contrast, the P2′ and P4′ substrate positions showed minimal activity. The influence of the P2 and P3 amino acids on ground state binding and the P4 position for transition state stabilization was identified through single substrate kinetics with optimal and suboptimal substrate sequences. The specificities observed for dengue NS2B/NS3 have features in common with the physiological cleavage sites in the dengue polyprotein; however, all sites reveal previously unrecognized suboptimal sequences.
登录
查看更多内容
影响因子：
3.7
作者：
ARIAS, CF;PREUGSCHAT, F;STRAUSS, JH
通讯作者：
STRAUSS, JH
登革热 2 病毒非结构蛋白 NS4A 和 NS4B 的体外和体内加工。
影响因子：
3.7
作者：
Preugschat,F;Strauss,JH
通讯作者：
Strauss,JH
影响因子：
56.9
作者：
HEDSTROM, L;SZILAGYI, L;RUTTER, WJ
通讯作者：
RUTTER, WJ
DOI：
10.1073/pnas.140132697
发表时间：
2000-07-05
影响因子：
11.1
作者：
Harris, JL;Backes, BJ;Craik, CS
通讯作者：
Craik, CS