1PM6

Solution Structure of Full-Length Excisionase (Xis) from Bacteriophage HK022

1PM6 の概要

• エントリーDOI: 10.2210/pdb1pm6/pdb
• 分子名称: Excisionase (1 entity in total)
• 機能のキーワード: antiparallel beta-sheet, winged-helix, cis-trans-trans triproline, gene regulation
• 由来する生物種: Enterobacteria phage HK022
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 8638.06

構造登録者

Rogov, V.V.,Luecke, C.,Muresanu, L.,Wienk, H.,Kleinhaus, I.,Werner, K.,Loehr, F.,Pristovsek, P.,Rueterjans, H. (登録日: 2003-06-10, 公開日: 2003-12-30, 最終更新日: 2024-05-22)

主引用文献

Rogov, V.V.,Lucke, C.,Muresanu, L.,Wienk, H.,Kleinhaus, I.,Werner, K.,Lohr, F.,Pristovsek, P.,Ruterjans, H.
Solution structure and stability of the full-length excisionase from bacteriophage HK022.
Eur.J.Biochem., 270:4846-4858, 2003

PubMed Abstract: Heteronuclear high-resolution NMR spectroscopy was employed to determine the solution structure of the excisionase protein (Xis) from the lambda-like bacteriophage HK022 and to study its sequence-specific DNA interaction. As wild-type Xis was previously characterized as a generally unstable protein, a biologically active HK022 Xis mutant with a single amino acid substitution Cys28-->Ser was used in this work. This substitution has been shown to diminish the irreversibility of Xis denaturation and subsequent degradation, but does not affect the structural or thermodynamic properties of the protein, as evidenced by NMR and differential scanning calorimetry. The solution structure of HK022 Xis forms a compact, highly ordered protein core with two well-defined alpha-helices (residues 5-11 and 18-27) and five beta-strands (residues 2-4, 30-31, 35-36, 41-44 and 48-49). These data correlate well with 1H2O-2H2O exchange experiments and imply a different organization of the HK022 Xis secondary structure elements in comparison with the previously determined structure of the bacteriophage lambda excisionase. Superposition of both Xis structures indicates a better correspondence of the full-length HK022 Xis to the typical 'winged-helix' DNA-binding motif, as found, for example, in the DNA-binding domain of the Mu-phage repressor. Residues 51-72, which were not resolved in the lambda Xis, do not show any regular structure in HK022 Xis and thus appear to be completely disordered in solution. The resonance assignments have shown, however, that an unusual connectivity exists between residues Asn66 and Gly67 owing to asparagine-isoaspartyl isomerization. Such an isomerization has been previously observed and characterized only in eukaryotic proteins.

PubMed: 14653811
DOI: 10.1111/j.1432-1033.2003.03884.x

実験手法

SOLUTION NMR