Title
NMR Structure of the N-terminal Coiled Coil Domain of the Andes Hantavirus Nucleocapsid Protein
Author
Yu Wang, Daniel M. Boudreaux, D. Fernando Estrada, Chet W. Egan, Stephen C. St. Jeor, Roberto N. De Guzman
Year
2008
Journal
The Journal of Biological Chemistry
Abstract
The hantaviruses are emerging infectious viruses that in humans can cause a cardiopulmonary syndrome or a hemorrhagic fever with renal syndrome. The nucleocapsid (N) is the most abundant viral protein, and during viral assembly, the N protein forms trimers and packages the viral RNA genome. Here, we report the NMR structure of the N-terminal domain (residues 1–74, called N1–74) of the Andes hantavirus N protein. N1–74 forms two long helices (α1 and α2) that intertwine into a coiled coil domain. The conserved hydrophobic residues at the helix α1-α2 interface stabilize the coiled coil; however, there are many conserved surface residues whose function is not known. Site-directed mutagenesis, CD spectroscopy, and immunocytochemistry reveal that a point mutation in the conserved basic surface formed by Arg22 or Lys26 lead to antibody recognition based on the subcellular localization of the N protein. Thus, Arg22 and Lys26 are likely involved in a conformational change or molecular recognition when the N protein is trafficked from the cytoplasm to the Golgi, the site of viral assembly and maturation.
Full Article
Instrument
J-815
Keywords
Circular dichroism, Secondary structure, Protein denaturation, Thermal stability, Biochemistry