Journal article
Expression of Functional Soluble Human α-Globin Chains of Hemoglobin in Bacteria
Protein expression and purification, v 20(1)
Oct 2000
PMID: 11035948
Featured in Collection : UN Sustainable Development Goals @ Drexel
Abstract
Individual, soluble human α-globin chains were expressed in bacteria with exogenous heme and methionine aminopeptidase. The yields of soluble α chains in bacteria were comparable to those of recombinant non-α chains expressed under the same conditions. Molecular mass and gel-filtration properties of purified recombinant α chains were the same as those of authentic human α chains. Biochemical and biophysical properties of isolated α chains were identical to those of native human α chains as assessed by UV/vis, circular dichroism (CD), and nuclear magnetic resonance (NMR) spectroscopy which contrasts with previous results of refolded precipitated α chains made in the presence of heme in vitro (M. T. Sanna et al., J. Biol. Chem. 272, 3478–3486, 1997). Mixtures of purified, soluble recombinant α-globin and native β-globin chains formed heterotetramers in vitro, and oxygen- and CO-binding properties as well as the heme environment of the assembled tetramers were experimentally indistinguishable from those of native human Hb A. UV/vis, CD, and NMR spectra of assembled Hb A were also the same as those of human Hb A. These results indicate that individual expressed α chains are stable in bacteria and fold properly in vivo and that they then can assemble with free β chains to form hemoglobin heterotetramers in vivo as well as in vitro.
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Details
- Title
- Expression of Functional Soluble Human α-Globin Chains of Hemoglobin in Bacteria
- Creators
- Kazuhiko Adachi - Children's Hospital of PhiladelphiaTakamasa Yamaguchi - Children's Hospital of PhiladelphiaYi Yang - Children's Hospital of PhiladelphiaPatrick T. Konitzer - Children's Hospital of PhiladelphiaJian Pang - Children's Hospital of PhiladelphiaKonda S. Reddy - University of PennsylvaniaMaria Ivanova - Drexel UniversityFrank Ferrone - Drexel UniversitySaul Surrey - Dupont HospitalYonghong F Yang - Psychiatry
- Publication Details
- Protein expression and purification, v 20(1)
- Publisher
- Elsevier
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- Psychiatry; Physics
- Web of Science ID
- WOS:000089916800005
- Scopus ID
- 2-s2.0-0033810198
- Other Identifier
- 991019167691604721
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- Collaboration types
- Domestic collaboration
- Web of Science research areas
- Biochemical Research Methods
- Biochemistry & Molecular Biology
- Biotechnology & Applied Microbiology