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Title
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Efficient expression, purification and crystallisation of two hyperthermostable enzymes of histidine biosynthesis.
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Authors
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R.Thoma,
G.Obmolova,
D.A.Lang,
M.Schwander,
P.Jenö,
R.Sterner,
M.Wilmanns.
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Ref.
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FEBS Lett, 1999,
454,
1-6.
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PubMed id
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Abstract
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Enzymes from hyperthermophiles can be efficiently purified after expression in
mesophilic hosts and are well-suited for crystallisation attempts. Two enzymes
of histidine biosynthesis from Thermotoga maritima,
N'-((5'-phosphoribosyl)-formimino)-5-aminoimidazol-4-carb oxamid ribonucleotide
isomerase and the cyclase moiety of imidazoleglycerol phosphate synthase, were
overexpressed in Escherichia coli, both in their native and
seleno-methionine-labelled forms, purified by heat precipitation of host
proteins and crystallised.
N'-((5'-phosphoribosyl)-formimino)-5-aminoimidazol-4-carb oxamid ribonucleotide
isomerase crystallised in four different forms, all suitable for X-ray structure
solution, and the cyclase moiety of imidazoleglycerol phosphate synthase yielded
one crystal form that diffracted to atomic resolution. The obtained crystals
will enable the determination of the first three-dimensional structures of
enzymes from the histidine biosynthetic pathway.
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