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Structural, functional, and genetic characterization of Gastrophilus hemoglobin

Sylvia Dewilde, Mark Blaxter, Marie-Louise Van Hauwaert, Koen Van Houte, Alessandra Pesce, Nathalie Griffon, Laurent Kiger, Michael C Marden, Sven Vermeire, Jacques Vanfleteren UGent, et al. (1998) JOURNAL OF BIOLOGICAL CHEMISTRY. 273(49). p.32467-32474
abstract
Hemoglobin of Gastrophilus intestinalis (Insecta, Diptera), was purified and characterized. At least two isoforms have been identified by isoelectrofocusing, mass spectrometry, and genomic Southern blotting. Functional studies show a high oxygen affinity due to a low ligand dissociation rate (k(off) = 2.4 s(-1)) and a relatively high autoxidation rate (t(1/2) = 1.6/h), The globins were separated under denaturing conditions, and the sequence of Hb1 (M-r = 17,965 +/- 2) was determined at the protein and DNA level, The open reading frame codes for a polypeptide of 150 amino acids. Although the globin is distantly related to globins from other species, it has a low penalty score against globin templates. Freshly isolated hemoglobin was crystallized from polyethylene glycol. Crystals contain two hemoglobin molecules per asymmetric unit. Solution of the three-dimensional structure by molecular replacement could not be achieved, possibly due to the presence of three protein isoforms in the crystals. In order to determine its three-dimensional structure, G. intestinalis Hb1 was overexpressed in Escherichia coli, resulting in a fully functional molecule as confirmed by Ligand binding affinity. The globin gene contains two introns at positions D7.0 and G7.0. The D7.0 intron is unprecedented, suggesting that globin gene evolution is much more complex than originally thought.
Please use this url to cite or link to this publication:
author
organization
year
type
journalArticle (original)
publication status
published
subject
keyword
INSECT CHIRONOMUS-THUMMI, NONVERTEBRATE GLOBIN GENES, AMINO-ACID SUBSTITUTION, SPERM WHALE MYOGLOBIN, LIGAND-BINDING, ASCARIS HEMOGLOBIN, INTRON POSITIONS, CENTRAL EXON, EVOLUTION, SEQUENCES
journal title
JOURNAL OF BIOLOGICAL CHEMISTRY
J. Biol. Chem.
volume
273
issue
49
pages
32467 - 32474
Web of Science type
Article
Web of Science id
000077329100017
ISSN
0021-9258
DOI
10.1074/jbc.273.49.32467
language
English
UGent publication?
yes
classification
A1
copyright statement
I have transferred the copyright for this publication to the publisher
id
181007
handle
http://hdl.handle.net/1854/LU-181007
date created
2004-01-14 13:41:00
date last changed
2016-12-19 15:38:46
@article{181007,
  abstract     = {Hemoglobin of Gastrophilus intestinalis (Insecta, Diptera), was purified and characterized. At least two isoforms have been identified by isoelectrofocusing, mass spectrometry, and genomic Southern blotting. Functional studies show a high oxygen affinity due to a low ligand dissociation rate (k(off) = 2.4 s(-1)) and a relatively high autoxidation rate (t(1/2) = 1.6/h), The globins were separated under denaturing conditions, and the sequence of Hb1 (M-r = 17,965 +/- 2) was determined at the protein and DNA level, The open reading frame codes for a polypeptide of 150 amino acids. Although the globin is distantly related to globins from other species, it has a low penalty score against globin templates. 
Freshly isolated hemoglobin was crystallized from polyethylene glycol. Crystals contain two hemoglobin molecules per asymmetric unit. Solution of the three-dimensional structure by molecular replacement could not be achieved, possibly due to the presence of three protein isoforms in the crystals. In order to determine its three-dimensional structure, G. intestinalis Hb1 was overexpressed in Escherichia coli, resulting in a fully functional molecule as confirmed by Ligand binding affinity. 
The globin gene contains two introns at positions D7.0 and G7.0. The D7.0 intron is unprecedented, suggesting that globin gene evolution is much more complex than originally thought.},
  author       = {Dewilde, Sylvia and Blaxter, Mark and Van Hauwaert, Marie-Louise and Van Houte, Koen and Pesce, Alessandra and Griffon, Nathalie and Kiger, Laurent and Marden, Michael C and Vermeire, Sven and Vanfleteren, Jacques and Esmans, Eddy and Moens, Luc},
  issn         = {0021-9258},
  journal      = {JOURNAL OF BIOLOGICAL CHEMISTRY},
  keyword      = {INSECT CHIRONOMUS-THUMMI,NONVERTEBRATE GLOBIN GENES,AMINO-ACID SUBSTITUTION,SPERM WHALE MYOGLOBIN,LIGAND-BINDING,ASCARIS HEMOGLOBIN,INTRON POSITIONS,CENTRAL EXON,EVOLUTION,SEQUENCES},
  language     = {eng},
  number       = {49},
  pages        = {32467--32474},
  title        = {Structural, functional, and genetic characterization of Gastrophilus hemoglobin},
  url          = {http://dx.doi.org/10.1074/jbc.273.49.32467},
  volume       = {273},
  year         = {1998},
}

Chicago
Dewilde, Sylvia, Mark Blaxter, Marie-Louise Van Hauwaert, Koen Van Houte, Alessandra Pesce, Nathalie Griffon, Laurent Kiger, et al. 1998. “Structural, Functional, and Genetic Characterization of Gastrophilus Hemoglobin.” Journal of Biological Chemistry 273 (49): 32467–32474.
APA
Dewilde, S., Blaxter, M., Van Hauwaert, M.-L., Van Houte, K., Pesce, A., Griffon, N., Kiger, L., et al. (1998). Structural, functional, and genetic characterization of Gastrophilus hemoglobin. JOURNAL OF BIOLOGICAL CHEMISTRY, 273(49), 32467–32474.
Vancouver
1.
Dewilde S, Blaxter M, Van Hauwaert M-L, Van Houte K, Pesce A, Griffon N, et al. Structural, functional, and genetic characterization of Gastrophilus hemoglobin. JOURNAL OF BIOLOGICAL CHEMISTRY. 1998;273(49):32467–74.
MLA
Dewilde, Sylvia, Mark Blaxter, Marie-Louise Van Hauwaert, et al. “Structural, Functional, and Genetic Characterization of Gastrophilus Hemoglobin.” JOURNAL OF BIOLOGICAL CHEMISTRY 273.49 (1998): 32467–32474. Print.