4J2A

RB69 DNA Polymerase L415A Ternary Complex

4J2A の概要

• エントリーDOI: 10.2210/pdb4j2a/pdb
• 関連するPDBエントリー: 4J2B 4J2D 4J2E
• 分子名称: DNA polymerase, DNA (5'-D(*TP*CP*GP*AP*GP*TP*AP*AP*GP*CP*AP*GP*TP*CP*CP*GP*CP*G)-3'), DNA (5'-D(*GP*CP*GP*GP*AP*CP*TP*GP*CP*TP*TP*AP*C)-3'), ... (6 entities in total)
• 機能のキーワード: rb69, dna polymerase, l415a, polymerase, transferase-dna complex, transferase/dna
• 由来する生物種: Enterobacteria phage RB69
• タンパク質・核酸の鎖数: 3
• 化学式量合計: 114622.70

構造登録者

Xia, S.,Wang, J.,Konigsberg, W.H. (登録日: 2013-02-04, 公開日: 2014-02-19, 最終更新日: 2024-02-28)

主引用文献

Xia, S.,Wood, M.,Bradley, M.J.,De La Cruz, E.M.,Konigsberg, W.H.
Alteration in the cavity size adjacent to the active site of RB69 DNA polymerase changes its conformational dynamics.
Nucleic Acids Res., 41:9077-9089, 2013

PubMed Abstract: Internal cavities are a common feature of many proteins, often having profound effects on the dynamics of their interactions with substrate and binding partners. RB69 DNA polymerase (pol) has a hydrophobic cavity right below the nucleotide binding pocket at the tip of highly conserved L415 side chain. Replacement of this residue with Gly or Met in other B family pols resulted in higher mutation rates. When similar substitutions for L415 were introduced into RB69pol, only L415A and L415G had dramatic effects on pre-steady-state kinetic parameters, reducing base selectivity by several hundred fold. On the other hand, the L415M variant behaved like the wild-type. Using a novel tC(o)-tCnitro Förster Resonance Energy Transfer (FRET) assay, we were able to show that the partition of the primer terminus between pol and exonuclease (exo) domains was compromised with the L415A and L415G mutants, but not with the L415M variant. These results could be rationalized by changes in their structures as determined by high resolution X-ray crystallography.

PubMed: 23921641
DOI: 10.1093/nar/gkt674

実験手法

X-RAY DIFFRACTION (1.8 Å)