TY - JOUR
T1 - Recombinant human hemoglobin
T2 - Expression and refolding of β-globin from Escherichia coli
AU - Fronticelli, Clara
AU - O'Donnell, J. Kevin
AU - Brinigar, William S.
PY - 1991/10
Y1 - 1991/10
N2 - 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.
AB - 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.
KW - Hemoglobin
KW - protein folding
KW - recombinant hemoglobin
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U2 - 10.1007/BF01025477
DO - 10.1007/BF01025477
M3 - Article
C2 - 1799407
AN - SCOPUS:0026002149
SN - 0277-8033
VL - 10
SP - 495
EP - 501
JO - Journal of Protein Chemistry
JF - Journal of Protein Chemistry
IS - 5
ER -