Abstract
The structured core of the N-terminal 3'-5' exonuclease domain of epsilon, the proofreading subunit of Escherichia coli DNA polymerase III, was defined by multidimensional NMR experiments with uniformly (15)N-labeled protein: it comprises residues between Ile-4 and Gln-181. A 185-residue fragment, termed epsilon(1-185), was crystallized by the hanging drop vapor diffusion method in the presence of thymidine-5'-monophosphate, a product inhibitor, and Mn(2+) at pH 5.8. The crystals are tetragonal, with typical dimensions 0.2 mm x 0.2 mm x 1.0 mm, grow over about 2 weeks at 4 degrees C, and diffract X-rays to 2.0 A. The space group was determined to be P4(n)2(1)2 (n = 0, 1, 2, 3), with unit cell dimensions a = 60.8 A, c = 111.4 A.
Copyright 2000 Academic Press.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Catalytic Domain*
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Chymotrypsin / metabolism
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Crystallization
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Crystallography, X-Ray
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DNA Polymerase III / chemistry*
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DNA Polymerase III / genetics
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DNA Polymerase III / metabolism
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Electrophoresis, Polyacrylamide Gel
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Escherichia coli / enzymology*
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Exodeoxyribonucleases / chemistry*
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Exodeoxyribonucleases / genetics
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Exodeoxyribonucleases / metabolism
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Manganese / metabolism
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Mutagenesis
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Nuclear Magnetic Resonance, Biomolecular
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Peptide Fragments / chemistry
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Peptide Fragments / metabolism
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Protein Structure, Tertiary
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Protein Subunits
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Sequence Alignment
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Thymidine Monophosphate / metabolism
Substances
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Peptide Fragments
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Protein Subunits
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Thymidine Monophosphate
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Manganese
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DNA Polymerase III
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Exodeoxyribonucleases
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Chymotrypsin