1vsd
|
ASV INTEGRASE CORE DOMAIN WITH MG(II) COFACTOR AND HEPES LIGAND, HIGH MG CONCENTRATION FORM
OverviewOverview
BACKGROUND: Members of the structurally-related superfamily of enzymes, that includes RNase H, RuvC resolvase, MuA transposase, and retroviral, integrase require divalent cations for enzymatic activity. So far, cation, positions are reported in the X-ray crystal structures of only two of, these proteins, E. coli and human immunodeficiency virus 1 (HIV-1) RNase, H. Details of the placement of metal ions in the active site of retroviral, integrases are necessary for the understanding of the catalytic mechanism, of these enzymes. RESULTS: The structure of the enzymatically active, catalytic domain (residues 52-207) of avian sarcoma virus integrase (ASV, IN) has been solved in the presence of divalent cations (Mn2+ or Mg2+), at, 1.7-2.2 A resolution. A single ion of either type interacts with the, carboxylate groups of the active site aspartates and uses four water, molecules to complete its octahedral coordination. The placement of the, aspartate side chains and metal ions is very similar to that observed in, the RNase H members of this superfamily; however, the conformation of the, catalytic aspartates in the active site of ASV IN differs significantly, from that reported for the analogous residues in HIV-1 IN. CONCLUSIONS:, Binding of the required metal ions does not lead to significant structural, modifications in the active site of the catalytic domain of ASV IN. This, indicates that at least one metal-binding site is preformed in the, structure, and suggests that the observed constellation of the acidic, residues represents a catalytically competent active site. Only a single, divalent cation was observed even at extremely high concentrations of the, metals. We conclude that either only one metal ion is needed for, catalysis, or that a second metal-binding site can only exist in the, presence of substrate and/or other domains of the protein. The unexpected, differences between the active sites of ASV IN and HIV-1 IN remain, unexplained; they may reflect the effects of crystal contacts on the, active site of HIV-1 IN, or a tendency for structural polymorphism.
About this StructureAbout this Structure
1VSD is a Single protein structure of sequence from Rous sarcoma virus with MG, OHE and EPE as ligands. Full crystallographic information is available from OCA.
ReferenceReference
The catalytic domain of avian sarcoma virus integrase: conformation of the active-site residues in the presence of divalent cations., Bujacz G, Jaskolski M, Alexandratos J, Wlodawer A, Merkel G, Katz RA, Skalka AM, Structure. 1996 Jan 15;4(1):89-96. PMID:8805516
Page seeded by OCA on Wed Nov 21 05:07:22 2007