Publication details
Using cryoEM Reconstruction and Phase Extension to Determine Crystal Structure of Bacteriophage I center dot 6 Major Capsid Protein
| Authors | |
|---|---|
| Year of publication | 2013 |
| Type | Article in Periodical |
| Magazine / Source | The Protein Journal |
| MU Faculty or unit | |
| Citation | |
| Web | http://link.springer.com/article/10.1007%2Fs10930-013-9526-x |
| Doi | http://dx.doi.org/10.1007/s10930-013-9526-x |
| Field | Biochemistry |
| Keywords | Molecular replacement; Cryo-electron microscopy; Non-crystallographic symmetry; Virus capsid protein; Phase extension |
| Description | Bacteriophage I center dot 6 is a double-stranded RNA virus that has been extensively studied as a model organism. Here we describe structure determination of I center dot 6 major capsid protein P1. The protein crystallized in base centered orthorhombic space group C222(1). Matthews's coefficient indicated that the crystals contain from four to seven P1 subunits in the crystallographic asymmetric unit. The self-rotation function had shown presence of fivefold axes of non-crystallographic symmetry in the crystals. Thus, electron density map corresponding to a P1 pentamer was excised from a previously determined cryoEM reconstruction of the I center dot 6 procapsid at 7 resolution and used as a model for molecular replacement. The phases for reflections at higher than 7 resolution were obtained by phase extension employing the fivefold non-crystallographic symmetry present in the crystal. The averaged 3.6 -resolution electron density map was of sufficient quality to allow model building. |