Crystallization and preliminary X-ray analysis of the dsDNA bacteriophage HK97 mature empty capsid

Virology
W R WikoffJohn E Johnson

Abstract

HK97 is a temperate dsDNA bacteriophage of Escherichia coli that is structurally similar to phage lambda, with an icosahedral head of triangulation (7) number 7. Although the capsids of several large dsDNA phages have been studied extensively using a variety of biophysical approaches, no high-resolution structure is available. We have grown crystals of mature but empty bacteriophage HK97 capsids that diffract to at least 3.5 A using synchrotron radiation. The HK97 Head II crystals are the first capsid crystals from a dsDNA bacteriophage that diffract X-rays to high resolution. A capsid precursor (prohead) was made in vivo by expressing capsid proteins in E. coli. This prohead was purified, converted to Head II in vitro, and used to grow crystals. The empty Head II has the same form as the mature HK97 capsid, but without DNA. The crystals were grown in a mixture of ammonium sulfate and PEG 8000, directly in an X-ray capillary to minimize crystal handling. The unit cell is monoclinic, with dimensions a = 580 A, b = 625 A, c = 790 A, beta = 90.0 degrees and two particles per unit cell.

References

Oct 13, 1995·Journal of Molecular Biology·J F ConwayA C Steven

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Citations

Apr 22, 2008·Quarterly Reviews of Biophysics·Elmar NurmemmedovAlex Evilevitch
Dec 4, 2003·Journal of Molecular Biology·Charlotte HelgstrandLars Liljas
Jan 16, 1999·Current Biology : CB·J E Johnson, W R Wikoff
May 5, 1999·Current Biology : CB·W R Wikoff, J E Johnson
Nov 3, 2015·Journal of Synchrotron Radiation·Karolina MichalskaAndrzej Joachimiak
Mar 11, 2003·Microbiology and Molecular Biology Reviews : MMBR·Eric S MillerWolfgang Rüger
Oct 26, 1999·Journal of Molecular Biology·M F Moody

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