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Bacteriophage N4 large terminase: expression, purification and X-ray crystallographic analysis.


ABSTRACT: Genome packaging is a critical step in the assembly of dsDNA bacteriophages and is carried out by a powerful molecular motor known as the large terminase. To date, wild-type structures of only two large terminase proteins are available, and more structural information is needed to understand the genome-packaging mechanism. Towards this goal, the large and small terminase proteins from bacteriophage N4, which infects the Escherichia coli K12 strain, have been cloned, expressed and purified. The purified putative large terminase protein hydrolyzes ATP, and this is enhanced in the presence of the small terminase. The large terminase protein was crystallized using the sitting-drop vapour-diffusion method and the crystal diffracted to 2.8?Å resolution using a home X-ray source. Analysis of the X-ray diffraction data showed that the crystal belonged to space group P212121, with unit-cell parameters a = 53.7, b = 93.6, c = 124.9?Å, ? = ? = ? = 90°. The crystal had a solvent content of 50.2% and contained one molecule in the asymmetric unit.

SUBMITTER: Wangchuk J 

PROVIDER: S-EPMC5894105 | biostudies-literature | 2018 Apr

REPOSITORIES: biostudies-literature

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Bacteriophage N4 large terminase: expression, purification and X-ray crystallographic analysis.

Wangchuk Jigme J   Prakash Prem P   Bhaumik Prasenjit P   Kondabagil Kiran K  

Acta crystallographica. Section F, Structural biology communications 20180322 Pt 4


Genome packaging is a critical step in the assembly of dsDNA bacteriophages and is carried out by a powerful molecular motor known as the large terminase. To date, wild-type structures of only two large terminase proteins are available, and more structural information is needed to understand the genome-packaging mechanism. Towards this goal, the large and small terminase proteins from bacteriophage N4, which infects the Escherichia coli K12 strain, have been cloned, expressed and purified. The p  ...[more]

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