Abstract

Hemoglobins under oxidative stress aggregate to give Heinz bodies that become attached to the cell membrane. Membrane distortion and oxidative membrane damage result in lysis of the erythrocyte. We have found that the reaction of hemoglobin with arylhydrazines gives a complex with the aryl group directly bound to the prosthetic heme iron atom and, under certain conditions, results in covalent crosslinking of the proteins. The present project is intended to (a) elucidate the structure and chemistry of the iron-aryl complexes, (b) determine if the complexes can be transferred to membranes, (c) determine if Heinz body formation can be monitored in intact erythrocytes by NMR, (d) explore the role of tyrosines proximal to the heme prosthetic groups in crosslinking, (d) investigate the mechanism and generality of direct lipid oxidation by hemoglobin, and (e) characterize sulfhemoglobin. The proposed work should provide important information not only on hemoprotein function and hemolytic mechanisms but should provide basic data relevant to the question of how damaged proteins are recognized for accelerated degradation.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK030297-06
Application #
3229367
Study Section
Hematology Subcommittee 2 (HEM)
Project Start
1982-01-01
Project End
1989-12-31
Budget Start
1987-01-01
Budget End
1987-12-31
Support Year
6
Fiscal Year
1987
Total Cost
Indirect Cost

  Institution

Name
University of California San Francisco
Department
Type
Schools of Pharmacy
DUNS #
073133571
City
San Francisco
State
CA
Country
United States
Zip Code
94143

  Publications

Peng, Dungeng; Ogura, Hiroshi; Ma, Li-Hua et al. (2013) Solution NMR characterization of magnetic/electronic properties of azide and cyanide-inhibited substrate complexes of human heme oxygenase: implications for steric ligand tilt. J Inorg Biochem 121:179-86
Meitzler, Jennifer L; Hinde, Sara; Bánfi, Botond et al. (2013) Conserved cysteine residues provide a protein-protein interaction surface in dual oxidase (DUOX) proteins. J Biol Chem 288:7147-57
Varfaj, Fatbardha; Lampe, Jed N; Ortiz de Montellano, Paul R (2012) Role of cysteine residues in heme binding to human heme oxygenase-2 elucidated by two-dimensional NMR spectroscopy. J Biol Chem 287:35181-91
Nishida, Clinton R; Ortiz de Montellano, Paul R (2011) Bioactivation of antituberculosis thioamide and thiourea prodrugs by bacterial and mammalian flavin monooxygenases. Chem Biol Interact 192:21-5
Meitzler, Jennifer L; Ortiz de Montellano, Paul R (2011) Structural stability and heme binding potential of the truncated human dual oxidase 2 (DUOX2) peroxidase domain. Arch Biochem Biophys 512:197-203
Jiang, Yongying; Trnka, Michael J; Medzihradszky, Katalin F et al. (2009) Covalent heme attachment to the protein in human heme oxygenase-1 with selenocysteine replacing the His25 proximal iron ligand. J Inorg Biochem 103:316-25
Peng, Dungeng; Ogura, Hiroshi; Zhu, Wenfeng et al. (2009) Coupling of the distal hydrogen bond network to the exogenous ligand in substrate-bound, resting state human heme oxygenase. Biochemistry 48:11231-42
Evans, John P; Kandel, Sylvie; Ortiz de Montellano, Paul R (2009) Isocyanides inhibit human heme oxygenases at the verdoheme stage. Biochemistry 48:8920-8
Ogura, Hiroshi; Evans, John P; Peng, Dungeng et al. (2009) The orbital ground state of the azide-substrate complex of human heme oxygenase is an indicator of distal H-bonding: implications for the enzyme mechanism. Biochemistry 48:3127-37
Meitzler, Jennifer L; Ortiz de Montellano, Paul R (2009) Caenorhabditis elegans and human dual oxidase 1 (DUOX1) ""peroxidase"" domains: insights into heme binding and catalytic activity. J Biol Chem 284:18634-43

Showing the most recent 10 out of 66 publications