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Title
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Design, construction, crystallization, and preliminary X-ray studies of a fine-tuning mutant (F133V) of module-substituted chimera hemoglobin.
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Authors
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T.Shirai,
M.Fujikake,
T.Yamane,
K.Inaba,
K.Ishimori,
I.Morishima.
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Ref.
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Proteins, 1998,
32,
263-267.
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PubMed id
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Abstract
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A chimera betaalpha-subunit of human hemoglobin was crystallized into a
carbonmonoxy form. The protein was assembled by substituting the structural
portion of a beta-subunit of hemoglobin (M4 module of the subunit) for its
counterpart in the alpha-subunit. In order to overcome the inherent instability
in the crystallization of the chimera subunit, a site-directed mutagenesis
(F133V) technique was employed based on a computer model. The crystal was used
for an X-ray diffraction study yielding a data set with a resolution of 2.5 A.
The crystal belongs to the monoclinic space group P21, with cell dimensions of a
= 62.9, b = 81.3, c = 55.1 A, and beta = 91.0 degrees . These dimensions are
similar to the crystallographic parameters of the native beta-subunit tetramers
in three different ligand states, one of which is a cyanide form that was also
crystallized in this study.
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