Crystallization and preliminary analysis of a dsDNA bacteriophage capsid intermediate: Prohead II of HK97

Crystallization and preliminary analysis of a dsDNA bacteriophage capsid intermediate: Prohead II... HK97 Prohead II is an early intermediate in the maturation of HK97, a T = 7 dsDNA‐tailed bacteriophage related to bacteriophage λ. Previously, selected capsid‐protein genes of HK97 were expressed in Escherichia coli and spontaneously assembled to form an icosahedral capsid that followed a maturation pathway closely similar to the authentic virion. The crystal structure of the mature HK97 capsid (Head II) made in this way was reported at 3.5 Å resolution. Additional high‐resolution structures of intermediates are needed to understand the maturation mechanism. The crystal structure of expressed Prohead II will elucidate the early steps of HK97 assembly. Crystals of the Prohead II mutant W336F were grown in 0.1 M HEPES pH 7.5, 0.2 M CaCl2 and 2–3% PEG 4000 at a Prohead II concentration of 16.5 mg ml−1. It was not possible to grow high‐quality crystals of wild‐type Prohead II. Diffraction was observed to 5 Å resolution from these crystals on beamline 14BM‐C at the Advanced Photon Source and data were collected to 5.5 Å with a completeness of 77%. The space group was P213, with unit‐cell parameter a = 707.0 Å and four particles in the unit cell. The particles are on the body diagonals of the cubic cell, with icosahedral threefold axes coincident with crystallographic threefold axes. Self‐rotation function and locked‐rotation function analysis determined the particle orientation and a one‐dimensional R‐­factor search along the body diagonal indicated that the particle centers were close to (1/4, 1/4, 1/4) and symmetry‐related positions. Molecular‐replacement averaging and phase extension are under way. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Acta Crystallographica Section D Wiley

Crystallization and preliminary analysis of a dsDNA bacteriophage capsid intermediate: Prohead II of HK97

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Publisher
Wiley
Copyright
Copyright © 2003 Wiley Subscription Services, Inc., A Wiley Company
ISSN
1399-0047
eISSN
1399-0047
DOI
10.1107/S0907444903022261
Publisher site
See Article on Publisher Site

Abstract

HK97 Prohead II is an early intermediate in the maturation of HK97, a T = 7 dsDNA‐tailed bacteriophage related to bacteriophage λ. Previously, selected capsid‐protein genes of HK97 were expressed in Escherichia coli and spontaneously assembled to form an icosahedral capsid that followed a maturation pathway closely similar to the authentic virion. The crystal structure of the mature HK97 capsid (Head II) made in this way was reported at 3.5 Å resolution. Additional high‐resolution structures of intermediates are needed to understand the maturation mechanism. The crystal structure of expressed Prohead II will elucidate the early steps of HK97 assembly. Crystals of the Prohead II mutant W336F were grown in 0.1 M HEPES pH 7.5, 0.2 M CaCl2 and 2–3% PEG 4000 at a Prohead II concentration of 16.5 mg ml−1. It was not possible to grow high‐quality crystals of wild‐type Prohead II. Diffraction was observed to 5 Å resolution from these crystals on beamline 14BM‐C at the Advanced Photon Source and data were collected to 5.5 Å with a completeness of 77%. The space group was P213, with unit‐cell parameter a = 707.0 Å and four particles in the unit cell. The particles are on the body diagonals of the cubic cell, with icosahedral threefold axes coincident with crystallographic threefold axes. Self‐rotation function and locked‐rotation function analysis determined the particle orientation and a one‐dimensional R‐­factor search along the body diagonal indicated that the particle centers were close to (1/4, 1/4, 1/4) and symmetry‐related positions. Molecular‐replacement averaging and phase extension are under way.

Journal

Acta Crystallographica Section DWiley

Published: Dec 1, 2003

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