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Title
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The structure of E.coli soluble inorganic pyrophosphatase at 2.7 A resolution.
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Authors
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J.Kankare,
G.S.Neal,
T.Salminen,
T.Glumoff,
T.].Glumhoff T [corrected to Glumoff,
B.S.Cooperman,
R.Lahti,
A.Goldman.
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Ref.
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Protein Eng, 1994,
7,
823-830.
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PubMed id
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Abstract
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The structure of E.coli soluble inorganic pyrophosphatase has been refined at
2.7 A 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 spreadout 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 motif that
seems to occur only in soluble inorganic pyrophosphatases.
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