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Design, construction, crystallization, and preliminary X‐ray studies of a fine‐tuning mutant (F133V) of module‐substituted chimera hemoglobin
Author(s) -
Shirai Tsuyoshi,
Fujikake Masahiro,
Yamane Takashi,
Inaba Kenji,
Ishimori Koichiro,
Morishima Isao
Publication year - 1998
Publication title -
proteins: structure, function, and bioinformatics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.699
H-Index - 191
eISSN - 1097-0134
pISSN - 0887-3585
DOI - 10.1002/(sici)1097-0134(19980815)32:3<263::aid-prot1>3.0.co;2-j
Subject(s) - protein subunit , crystallization , chimera (genetics) , crystallography , mutant , monoclinic crystal system , chemistry , hemoglobin , crystal structure , materials science , biochemistry , gene , organic chemistry
A chimera βα‐subunit of human hemoglobin was crystallized into a carbonmonoxy form. The protein was assembled by substituting the structural portion of a β‐subunit of hemoglobin (M4 module of the subunit) for its counterpart in the α‐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 Å. The crystal belongs to the monoclinic space group P 2 1 , with cell dimensions of a = 62.9, b = 81.3, c = 55.1 Å, and β = 91.0°. These dimensions are similar to the crystallographic parameters of the native β‐subunit tetramers in three different ligand states, one of which is a cyanide form that was also crystallized in this study. Proteins 32:263–267, 1998. © 1998 Wiley‐Liss, Inc.