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Title
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Production of crystallizable human chymase from a Bacillus subtilis system.
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Authors
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M.E.McGrath,
A.E.Osawa,
M.G.Barnes,
J.M.Clark,
K.D.Mortara,
B.F.Schmidt.
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Ref.
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Febs Lett, 1997,
413,
486-488.
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PubMed id
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Abstract
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A Bacillus subtilis strain deficient in seven extracellular proteases was used
to produce human mast cell chymase and is a viable expression system for serine
proteases and other classes of proteins. Chymase is produced at 0.3-0.5 mg/l and
is purified by three chromatography steps. Two crystal forms of PMSF-treated
chymase were optimized. The first is C2 with a=47.94 A, b=85.23 A, c=174.18 A,
beta=96.74 degrees, and diffracts to at least 2.1 A, while the second is
P212121, with cell dimensions a=43.93 A, b=58.16 A, and c=86.09 A, and a
diffraction limit of approximately 1.9 A. The first crystal form has either
three or four molecules/asymmetric unit, while the second has one
molecule/asymmetric unit.
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