THE STRUCTURE OF ESCHERICHIA-COLI SOLUBLE INORGANIC PYROPHOSPHATASE AT 2.7-ANGSTROM RESOLUTION

J KANKARE, GS NEAL, Tiina Salminen, T GLUMHOFF, BS COOPERMAN, R LAHTI, A GOLDMAN

    Forskningsoutput: TidskriftsbidragArtikelVetenskapligPeer review

    Sammanfattning

    The structure of E. coli soluble inorganic pyrophosphatase has been refined at 2.7 Angstrom resolution to an R-factor of 20.9%. The overall fold of the molecule is essentially the same as yeast pyrophosphatase, except that yeast pyrophosphatase is longer at both the N- and C-termini. Escherichia coli pyrophosphatase is a mixed alpha+beta protein with a complicated topology. The active site cavity, which is also very similar to the yeast enzyme, is formed by seven beta-strands and an alpha-helix and has a rather asymmetric distribution of charged residues. Our structure-based alignment extends and improves upon earlier sequence alignment studies; it shows that probably no more than 14, not 15-17 charged and polar residues are part of the conserved enzyme mechanism of pyrophosphatases. Six of these conserved residues, at the bottom of the active site cavity, form a tight group centred on Asp70 and probably bind the two essential Mg2+ ions. The others, more spread-out and more positively charged, presumably bind substrate. Escherichia coli pyrophosphatase has an extra aspartate residue in the active site cavity, which may explain why the two enzymes bind divalent cation differently. Based on the structure, we have identified a sequence moth that seems to occur only in soluble inorganic pyrophosphatases.
    OriginalspråkOdefinierat/okänt
    Sidor (från-till)823–830
    Antal sidor8
    TidskriftProtein Engineering
    Volym7
    Nummer7
    StatusPublicerad - 1994
    MoE-publikationstypA1 Tidskriftsartikel-refererad

    Nyckelord

    • SEQUENCE MOTIF
    • STRUCTURE COMPARISON
    • X-ray crystallography

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