Design, construction, crystallization, and preliminary X-ray studies of a fine-tuning mutant (F133V) of module-substituted chimera hemoglobin

Tsuyoshi Shirai, Masahiro Fujikake, Takashi Yamane, Kenji Inaba, Koichiro Ishimori, Isao Morishima

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

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 P21, 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.

Original languageEnglish
Pages (from-to)263-267
Number of pages5
JournalProteins: Structure, Function and Genetics
Volume32
Issue number3
DOIs
Publication statusPublished - 1998 Aug 15
Externally publishedYes

Keywords

  • Chimeric protein
  • Computer modeling
  • Hemoglobin H
  • Molecular evolution
  • Protein engineering

ASJC Scopus subject areas

  • Structural Biology
  • Biochemistry
  • Molecular Biology

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