Abstract

This core is responsible for the final stage in the preparation of synthetic mutants of human hemoglobin. There are three steps in obtaining a variant protein: The modification of the globin gene in the plasmid by site specific mutagenesis using the appropriate oligonucleotide the expression of this gene in E. coli, and the preparation of hemoglobin from the expressed protein. Obviously for any given variant the first step is done once while the subsequent steps are repeated as needed to produce adequate amounts of material. The first two steps will be carried out at the University of Iowa under Dr. Arnone's direction. The last step will be the responsibility of the Hemoglobin Assembly Core. Frozen cells will be shipped from the University of Iowa to this Core. As will be described, these cells will contain globin proteins on one of two forms. These proteins will be extracted from the cells and hemoglobin prepared which contain these globins. Synthetic mutants will be prepared which contain metalloporphyrins other than heme, or which contain a mixture of metalloporphyrins. Asymmetric hybrid hemoglobins will be prepared which either contain two different alphabeta dimers (mixed mutant hybrid hemoglobins) or both will be pared and stabilized by chemically cross-linking the hemoglobin tetramers.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Program Projects (P01)
Project #
5P01GM058890-02
Application #
6301800
Study Section
Project Start
2000-01-01
Project End
2000-12-31
Budget Start
1998-10-01
Budget End
1999-09-30
Support Year
2
Fiscal Year
2000
Total Cost
$209,639
Indirect Cost

  Institution

Name
University of Iowa
Department
Type
DUNS #
041294109
City
Iowa City
State
IA
Country
United States
Zip Code
52242

  Publications

Kwiatkowski, Laura D; Hui, Hilda L; Karasik, Ellen et al. (2007) Mutations of the betaN102 residue of HbA not only inhibit the ligand-linked T to Re state transition, but also profoundly affect the properties of the T state itself. Biochemistry 46:2037-49
Das, Tapan K; Dewilde, Sylvia; Friedman, Joel M et al. (2006) Multiple active site conformers in the carbon monoxide complexes of trematode hemoglobins. J Biol Chem 281:11471-9
Samuni, Uri; Roche, Camille J; Dantsker, David et al. (2006) Modulation of reactivity and conformation within the T-quaternary state of human hemoglobin: the combined use of mutagenesis and sol-gel encapsulation. Biochemistry 45:2820-35
Dantsker, David; Roche, Camille; Samuni, Uri et al. (2005) The position 68(E11) side chain in myoglobin regulates ligand capture, bond formation with heme iron, and internal movement into the xenon cavities. J Biol Chem 280:38740-55
Kavanaugh, Jeffrey S; Rogers, Paul H; Arnone, Arthur et al. (2005) Intersubunit interactions associated with Tyr42 alpha stabilize the quaternary-T tetramer but are not major quaternary constraints in deoxyhemoglobin. Biochemistry 44:3806-20
Kavanaugh, Jeffrey S; Rogers, Paul H; Arnone, Arthur (2005) Crystallographic evidence for a new ensemble of ligand-induced allosteric transitions in hemoglobin: the T-to-T(high) quaternary transitions. Biochemistry 44:6101-21
Dantsker, David; Samuni, Uri; Ouellet, Yannick et al. (2004) Viscosity-dependent relaxation significantly modulates the kinetics of CO recombination in the truncated hemoglobin TrHbN from Mycobacterium tuberculosis. J Biol Chem 279:38844-53
Tsuneshige, Antonio; Kanaori, Kenji; Samuni, Uri et al. (2004) Semihemoglobins, high oxygen affinity dimeric forms of human hemoglobin respond efficiently to allosteric effectors without forming tetramers. J Biol Chem 279:48959-67
Wheeler, Korin E; Lees, Nicholas S; Gurbiel, Ryszard J et al. (2004) Electrostatic influence on rotational mobilities of sol-gel-encapsulated solutes by NMR and EPR spectroscopies. J Am Chem Soc 126:13459-63
Chan, Nei-Li; Kavanaugh, Jeffrey S; Rogers, Paul H et al. (2004) Crystallographic analysis of the interaction of nitric oxide with quaternary-T human hemoglobin. Biochemistry 43:118-32

Showing the most recent 10 out of 37 publications