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Title
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Apomyoglobin as a molecular recognition surface: expression, reconstitution and crystallization of recombinant porcine myoglobin in Escherichia coli.
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Authors
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G.Dodson,
R.E.Hubbard,
T.J.Oldfield,
S.J.Smerdon,
A.J.Wilkinson.
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Ref.
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Protein Eng, 1988,
2,
233-237.
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PubMed id
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Abstract
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Recombinant porcine myoglobin has been produced in Escherichia coli using the
lambda cII fusion expression system of Nagai and Thøgersen [Nature, 309,
810-812 (1984)]. After processing and reconstitution with haem, the protein is
gel-electrophoretically and spectrophotometrically indistinguishable from native
pig myoglobin. Large crystals of both native and recombinant porcine myoglobin
were grown from 50 mM sodium phosphate, pH 7.1, 80% ammonium sulphate. The
crystals belong to space group C2 (a = 156.9 A, b = 42.0 A, c = 92.2 A, beta =
127.9 degrees) and diffract to a nominal 2.5 A resolution. We plan to explore
apomyoglobin as a binding surface in studies combining site-directed mutagenesis
and X-ray analysis. These experiments will be extended by studying the binding
of haem analogues to the mutant apoproteins.
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