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X‐Ray crystallographic studies of recombinant inorganic pyrophosphatase from Escherichia coli
Author(s) -
Oganessyan V.Yu.,
Kurilova S.A.,
Vorobyeva N.N.,
Nazarova T.I.,
Popov A.N.,
Lebedev A.A.,
Avaeva S.M.,
Harutyunyan E.H.
Publication year - 1994
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(94)00605-9
Subject(s) - inorganic pyrophosphatase , pyrophosphatase , crystallography , escherichia coli , chemistry , recombinant dna , molecule , enzyme , stereochemistry , biochemistry , organic chemistry , pyrophosphate , gene
An E. coli inorganic pyrophosphatase overproducer and a method for a large‐scale production of the homogeneous enzyme are described. The inorganic pyrophosphatase was crystallized in the form containing one subunit of a homohexameric molecule per asymmetric unit: space group R32, a = 110.4 Å, c = 76.8 Å. The electron density map to 2.5 Å resolution phased with Eu‐ and Hg‐derivatives (figure of merit, < m > = 0.51) was improved by the solvent flattening procedure (< m > = 0.77). The course of the polypeptide chain and the secondary structure elements, intersubunit contacts and positions of the active sites were characterized. Homology with S. cerevisiae inorganic pyrophosphatase structure was found.