Modulation of the affinity of the single‐stranded DNA‐binding protein of <i>Escherichia coli</i> (<i>E. coli</i> SSB) to poly(dT) by site‐directed mutagenesis
書誌事項
- 公開日
- 1989-02
- 権利情報
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- http://onlinelibrary.wiley.com/termsAndConditions#vor
- DOI
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- 10.1111/j.1432-1033.1989.tb14567.x
- 公開者
- Wiley
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説明
<jats:p>A vector for site‐directed mutagenesis and overproduction of the <jats:italic>Escherichia coli</jats:italic> single‐stranded‐DNA‐binding protein (<jats:italic>E. coli</jats:italic> SSB) was constructed. An <jats:italic>E. coli</jats:italic> strain carrying this vector produces up to 400 mg pure protein from 25 g wet cells. The vector was used to mutate specifically the Phe60 residue of <jats:italic>E. coli</jats:italic> SSB. Phe60 had been proposed to be located near the single‐stranded‐DNA‐binding site.</jats:p><jats:p>Substitution of the Phe60 residue by Val, Ser, Leu, His, Tyr and Trp gave proteins with no or only minor conformational changes, as detected by NMR spectroscopy.</jats:p><jats:p>The affinity of the mutant <jats:italic>E. coli</jats:italic> SSB proteins for single‐stranded DNA decreased in the order Trp > Phe (wild‐type) > Tyr > Leu > His > Val > Ser, leading to the conclusion that position 60 is a site of hydrophobic interaction of the protein with DNA.</jats:p>
収録刊行物
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- European Journal of Biochemistry
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European Journal of Biochemistry 179 (2), 399-404, 1989-02
Wiley