COMPUTATIONAL DESIGN OF PROTEINS TARGETING THE STEM REGION OF INFLUENZA

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 我们描述了一种通用的计算方法，用于设计结合目标大分子上的表面补丁的蛋白质。识别游离氨基酸残基和靶表面之间的有利相互作用，并用于锚重新设计的界面。该方法被用来设计结合1918年H1N1大流行病毒的流感血凝素（HA）茎上的保守表面补丁的蛋白质。亲和力成熟后，两个设计的蛋白质，HB 36和HB 80，结合H1和H5 HA低纳摩尔亲和力。此外，HB 80抑制在低pH值下诱导的HA融合构象变化。HB 36与1918/H1 HA复合的晶体结构显示，实际结合界面与计算设计模型中的界面几乎相同。这种设计的结合蛋白可能对诊断和治疗都有用。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. We describe a general computational method for designing proteins that bind a surface patch of interest on a target macromolecule. Favorable interactions between disembodied amino acid residues and the target surface are identified and used to anchor de novo designed interfaces. The method was used to design proteins that bind a conserved surface patch on the stem of the influenza hemagglutinin (HA) from the 1918 H1N1 pandemic virus. After affinity maturation, two of the designed proteins, HB36 and HB80, bind H1 and H5 HAs with low nanomolar affinity. Further, HB80 inhibits the HA fusogenic conformational changes induced at low pH. The crystal structure of HB36 in complex with 1918/H1 HA revealed that the actual binding interface is nearly identical to that in the computational design model. Such designed binding proteins may be useful for both diagnostics and therapeutics.