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Literature DB >> 1799407

Recombinant human hemoglobin: expression and refolding of beta-globin from Escherichia coli.

C Fronticelli¹, J K O'Donnell, W S Brinigar.

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-beta-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 beta-globin which can be folded in the presence of cyanohemin and native alpha-chains to produce a tetrameric hemoglobin with the functional properties of natural human hemoglobin.

Entities: Chemical Gene Species

Mesh：

Substances：

Year: 1991 PMID： 1799407 DOI： 10.1007/BF01025477

Source DB: PubMed Journal: J Protein Chem ISSN： 0277-8033

18 in total

1. A NEW METHOD FOR THE PREPARATION OF ALPHA AND BETA SUBUNITS OF HUMAN HEMOGLOBIN.

Authors: E BUCCI; C FRONTICELLI
Journal: J Biol Chem Date: 1965-01 Impact factor: 5.157

2. Characterization of haem disorder by circular dichroism.

Authors: H S Aojula; M T Wilson; A Drake
Journal: Biochem J Date: 1986-07-15 Impact factor: 3.857

3. On the determination of species fractions from ligand-binding data. Application to human hemoglobin.

Authors: E Di Cera; S J Gill
Journal: Biophys Chem Date: 1988-04 Impact factor: 2.352

4. Molecular weight analysis of oligopeptides by electrophoresis in polyacrylamide gel with sodium dodecyl sulfate.

Authors: R T Swank; K D Munkres
Journal: Anal Biochem Date: 1971-02 Impact factor: 3.365

5. Synthesis and sequence-specific proteolysis of hybrid proteins produced in Escherichia coli.

Authors: K Nagai; H C Thøgersen
Journal: Methods Enzymol Date: 1987 Impact factor: 1.600

6. Heme orientational disorder in reconstituted and native sperm whale myoglobin. Proton nuclear magnetic resonance characterizations by heme methyl deuterium labeling in the Met-cyano protein.

Authors: G N La Mar; N L Davis; D W Parish; K M Smith
Journal: J Mol Biol Date: 1983-08-25 Impact factor: 5.469

3. Cysteines beta93 and beta112 as probes of conformational and functional events at the human hemoglobin subunit interfaces.

Authors: G B Vásquez; M Karavitis; X Ji; I Pechik; W S Brinigar; G L Gilliland; C Fronticelli
Journal: Biophys J Date: 1999-01 Impact factor: 4.033

3 in total

Recombinant human hemoglobin: expression and refolding of beta-globin from Escherichia coli.

1. A NEW METHOD FOR THE PREPARATION OF ALPHA AND BETA SUBUNITS OF HUMAN HEMOGLOBIN.

2. Characterization of haem disorder by circular dichroism.

3. On the determination of species fractions from ligand-binding data. Application to human hemoglobin.

4. Molecular weight analysis of oligopeptides by electrophoresis in polyacrylamide gel with sodium dodecyl sulfate.

5. Synthesis and sequence-specific proteolysis of hybrid proteins produced in Escherichia coli.

6. Heme orientational disorder in reconstituted and native sperm whale myoglobin. Proton nuclear magnetic resonance characterizations by heme methyl deuterium labeling in the Met-cyano protein.

7. Membrane-covered thin-layer optical cell for gas-reaction studies of hemoglobin.

8. Oxygen binding constants for human hemoglobin tetramers.

9. Oxygen binding properties of human mutant hemoglobins synthesized in Escherichia coli.

10. Generation of beta-globin by sequence-specific proteolysis of a hybrid protein produced in Escherichia coli.

1. Two mutations in recombinant Hb beta F41(C7)Y, K82 (EF6)D show additive effects in decreasing oxygen affinity.

Review 2. Development of recombinant hemoglobin-based oxygen carriers.

3. Cysteines beta93 and beta112 as probes of conformational and functional events at the human hemoglobin subunit interfaces.