the presence of the iron-sulfur motif is important for the conformational stability of the antiviral protein, viperiniron-sulfur主题的存在是重要的抗病毒蛋白的构象稳定,viperin.pdfVIP

The Presence of the Iron-Sulfur Motif Is Important for the Conformational Stability of the Antiviral Protein, Viperin 1. 1. 1 2¤ Shubhasis Haldar , Simantasarani Paul , Nidhi Joshi , Anindya Dasgupta , Krishnananda Chattopadhyay1* 1 Structural Biology and Bioinformatics, Indian Institute of Chemical Biology, Council for Scientific and Industrial Research, Kolkata, West Bengal, India, 2 Infectious Diseases and Immunology Divisions, Indian Institute of Chemical Biology, Council for Scientific and Industrial Research, Kolkata, West Bengal, India Abstract Viperin, an antiviral protein, has been shown to contain a CX CX C motif, which is conserved in the radical S-adenosyl- 3 2 methionine (SAM) enzyme family. A triple mutant which replaces these three cysteines with alanines has been shown to have severe deficiency in antiviral activity. Since the crystal structure of Viperin is not available, we have used a combination of computational methods including multi-template homology modeling and molecular dynamics simulation to develop a low-resolution predicted structure. The results show that Viperin is an a -b protein containing iron-sulfur cluster at the center pocket. The calculations suggest that the removal of iron-sulfur cluster would lead to collapse of the protein tertiary structure. To verify these predictions, we have prepared, expressed and purified four mutant proteins. In three mutants individual cysteine residues were replaced by alanine residues while in the fourth all the cysteines were replaced by alanines. Conformational analyses using circul