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Characterisation of the DNA gyrase from the thermophilic eubacterium Thermus thermophilus.

Abstract：

:DNA gyrase is a type IIA topoisomerase found in bacteria but not in humans. The enzyme is required for bacterial DNA replication and transcription, and is an important antibacterial target that is sensitive to the widely-used fluoroquinolone drugs. Due to the emergence of fluoroquinolone resistance, the discovery of new classes of drugs that target DNA gyrase is urgent. The DNA gyrase holoenzyme is a heterodimer of subunit pairs (A2B2). The 90 kDa A subunits bind, cleave, and rejoin double stranded DNA. The enzyme introduces negative supercoils into closed circular bacterial DNA using ATP hydrolysis catalysed by the 70 kDa B subunits. Subdomains of DNA gyrase subunits have been crystallised for structural analysis and the resulting models used to improve drugs that target the DNA binding region and active site. While crystal structures are available for topoisomerase IV complexes with cleaved DNA, there is none for the complete DNA gyrase complex with substrate DNA bound. Thermophiles offer significant advantages in obtaining stable enzymes for structural and functional studies. In order to develop a capability for drug screening and structure-directed drug discovery we have reconstituted a functional and drug-sensitive DNA gyrase complex using heterologously expressed subunits from the thermophile Thermus thermophilus.

journal_name

Protein Expr Purif

journal_title

Protein expression and purification

authors

Aung HL,Samaranayaka CU,Enright R,Beggs KT,Monk BC

doi

10.1016/j.pep.2014.11.009

subject

Has Abstract

pub_date

2015-03-01 00:00:00

pages

62-7

eissn

1046-5928

issn

1096-0279

pii

S1046-5928(14)00270-8

journal_volume

107

pub_type

杂志文章

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