Production Of HIV And HIV Related Proteins For Structural Studies

Rev是HV-1的关键调控蛋白。它的功能是与病毒转录物结合，并影响未剪接mRNA的细胞核输出，从而允许结构蛋白的产生。尽管它很重要，但由于蛋白质聚集的趋势，其结构直到最近才被确定。抗体抗原结合域（Fab和scFv）可以介导难熔蛋白的共结晶。制备了蛋白质抗体复合物，产生晶体，并用x射线晶体学测定了Rev的结构。最近，Rev的高分辨率结构被用于模拟蛋白质-蛋白质相互作用或自我关联，这是Rev作用所必需的。此外，Rev与RNA和辅助蛋白的功能相互作用也正在研究中，目的是收集结构信息，这可能对靶向抗hiv干预有用。

Rev is a key regulatory protein of HV-1. Its function is to bind to viral transcripts and effect export from the nucleus of unspliced mRNA thereby allowing the production of structural proteins. Despite its importance, the structure has only been recently determined due to the tendency of the protein to aggregate. Antibody antigen binding domains (Fab and scFv) can mediate co-crystallization of refractory proteins. Protein antibody complexes were produced which generated crystals from which the structure of Rev was determined by X-ray crystallography. The more recent higher resolution structure of Rev is being used to model the protein-protein interactions or self-associations which are essential for Rev action. Also, the functional interactions of Rev with RNA and accessory proteins are being studied with the aim of gathering structural information which may be useful for targeted anti-HIV intervention. HIV protease, a homodimeric protein is essential in the viral life cycle and a major anti-HIV drug target. Peptides derived from the N- and C-terminal regions of the HIV-1 protease dimer interface inhibit protease activity by preventing dimerization (monomeric protein is inactive). In previous work, it was shown that the solubility and cell permeability of the peptides was enhanced by linking the transduction domain of HIV-Tat. A genetic construct was generated that can be used to study the first step in the cascade of HIV-1 protease Gag-Pro-Pol processing and show that reversible oxidation of cysteines can control the activity of the HIV-1 protease at this initial step. This assay system also has the potential for the screening of potential dimerization inhibitors.