Recombinant human hemoglobin: Expression and refolding of β-globin from Escherichia coli

Clara Fronticelli, J. Kevin O'Donnell, William S. Brinigar

Research output: Contribution to journalArticle

Abstract

A plasmid analogous to the one described by Nagai and Thogersen (Nature, 309, 810-812, 1984) has been constructed for the expression of globins in E. coli. Induction with nalidixic acid produces high yields of a fusion protein, NS1-FX-β-globin, where NS1 represents 81 residues of a flu virus protein and FX represents a blood-clotting Factor Xa recognition sequence, Ile-Glu-Gly-Arg. This fusion protein is readily solubilized in 50 mM NaOH and remains in solution when the pH is adjusted to 8.6. Under these conditions, the fusion protein is hydrolyzed by activated Factor X, giving authentic β-globin which can be folded in the presence of cyanohemin and native α-chains to produce a tetrameric hemoglobin with the functional properties of natural human hemoglobin.

Original languageEnglish (US)
Pages (from-to)495-501
Number of pages7
JournalJournal of Protein Chemistry
Volume10
Issue number5
DOIs
StatePublished - Oct 1991
Externally publishedYes

Fingerprint

Globins
Hemoglobin
Escherichia coli
Hemoglobins
Proteins
Factor Xa
Fusion reactions
Nalidixic Acid
Blood Coagulation Factors
Blood Coagulation
Viruses
Plasmids
Blood
Acids

Keywords

  • Hemoglobin
  • protein folding
  • recombinant hemoglobin

ASJC Scopus subject areas

  • Biochemistry

Cite this

Recombinant human hemoglobin : Expression and refolding of β-globin from Escherichia coli. / Fronticelli, Clara; O'Donnell, J. Kevin; Brinigar, William S.

In: Journal of Protein Chemistry, Vol. 10, No. 5, 10.1991, p. 495-501.

Research output: Contribution to journalArticle

Fronticelli, Clara ; O'Donnell, J. Kevin ; Brinigar, William S. / Recombinant human hemoglobin : Expression and refolding of β-globin from Escherichia coli. In: Journal of Protein Chemistry. 1991 ; Vol. 10, No. 5. pp. 495-501.
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