1VSM

ASV INTEGRASE CORE DOMAIN IN CITRATE BUFFER PH 5.0

1VSM の概要

• エントリーDOI: 10.2210/pdb1vsm/pdb
• 分子名称: PROTEIN (INTEGRASE) (2 entities in total)
• 機能のキーワード: endonuclease, transferase, hydrolase
• 由来する生物種: Rous sarcoma virus (strain Schmidt-Ruppin)
• 細胞内の位置: Matrix protein p19: Virion (Potential). Capsid protein p27: Virion (Potential). Nucleocapsid protein p12: Virion (Potential): P03354
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 15999.39

構造登録者

Lubkowski, J.,Yang, F.,Alexandratos, J.,Wlodawer, A. (登録日: 1998-09-18, 公開日: 1998-09-23, 最終更新日: 2023-08-23)

主引用文献

Lubkowski, J.,Yang, F.,Alexandratos, J.,Merkel, G.,Katz, R.A.,Gravuer, K.,Skalka, A.M.,Wlodawer, A.
Structural basis for inactivating mutations and pH-dependent activity of avian sarcoma virus integrase.
J.Biol.Chem., 273:32685-32689, 1998

PubMed Abstract: Crystallographic studies of the catalytic core domain of avian sarcoma virus integrase (ASV IN) have provided the most detailed picture so far of the active site of this enzyme, which belongs to an important class of targets for designing drugs against AIDS. Recently, crystals of an inactive D64N mutant were obtained under conditions identical to those used for the native enzyme. Data were collected at different pH values and in the presence of divalent cations. Data were also collected at low pH for the crystals of the native ASV IN core domain. In the structures of native ASV IN at pH 6.0 and below, as well as in all structures of the D64N mutants, the side chain of the active site residue Asx-64 (Asx denotes Asn or Asp) is rotated by approximately 150 degrees around the Calpha---Cbeta bond, compared with the structures at higher pH. In the new structures, this residue makes hydrogen bonds with the amide group of Asn-160, and thus, the usual metal-binding site, consisting of Asp-64, Asp-121, and Glu-157, is disrupted. Surprisingly, however, a single Zn2+ can still bind to Asp-121 in the mutant, without restoration of the activity of the enzyme. These structures have elucidated an unexpected mechanism of inactivation of the enzyme by lowering the pH or by mutation, in which a protonated side chain of Asx-64 changes its orientation and interaction partner.

PubMed: 9830010
DOI: 10.1074/jbc.273.49.32685

実験手法

X-RAY DIFFRACTION (2.15 Å)