GAPDH regulates cellular heme insertion into inducible nitric oxide synthase
- 4 October 2010
- journal article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences of the United States of America
- Vol. 107 (42), 18004-18009
- https://doi.org/10.1073/pnas.1008133107
Abstract
Heme proteins play essential roles in biology, but little is known about heme transport inside mammalian cells or how heme is inserted into soluble proteins. We recently found that nitric oxide (NO) blocks cells from inserting heme into several proteins, including cytochrome P450s, hemoglobin, NO synthases, and catalase. This finding led us to explore the basis for NO inhibition and to identify cytosolic proteins that may be involved, using inducible NO synthase (iNOS) as a model target. Surprisingly, we found that GAPDH plays a key role. GAPDH was associated with iNOS in cells. Pure GAPDH bound tightly to heme or to iNOS in an NO-sensitive manner. GAPDH knockdown inhibited heme insertion into iNOS and a GAPDH mutant with defective heme binding acted as a dominant negative inhibitor of iNOS heme insertion. Exposing cells to NO either from a chemical donor or by iNOS induction caused GAPDH to become S-nitrosylated at Cys152. Expressing a GAPDH C152S mutant in cells or providing a drug to selectively block GAPDH S-nitrosylation both made heme insertion into iNOS resistant to the NO inhibition. We propose that GAPDH delivers heme to iNOS through a process that is regulated by its S-nitrosylation. Our findings may uncover a fundamental step in intracellular heme trafficking, and reveal a mechanism whereby NO can govern the process.Keywords
This publication has 38 references indexed in Scilit:
- Nitric oxide blocks cellular heme insertion into a broad range of heme proteinsFree Radical Biology & Medicine, 2010
- Novel roles for GAPDH in cell death and carcinogenesisCell Death & Differentiation, 2009
- Trafficking of Heme and Porphyrins in MetazoaChemical Reviews, 2009
- Protein S-nitrosylation in health and disease: a current perspectiveTrends in Molecular Medicine, 2009
- Negative feedback maintenance of heme homeostasis by its receptor, Rev-erbαGenes & Development, 2009
- The Hsp90 Chaperone Machinery Regulates Signaling by Modulating Ligand Binding CleftsPublished by Elsevier BV ,2008
- Regulated Protein Denitrosylation by Cytosolic and Mitochondrial ThioredoxinsScience, 2008
- Heme: a versatile signaling molecule controlling the activities of diverse regulators ranging from transcription factors to MAP kinasesCell Research, 2006
- Nitric Oxide–GAPDH–Siah: A Novel Cell Death CascadeCellular and Molecular Neurobiology, 2006
- Succinylacetone, a potent inhibitor of heme biosynthesis: Effect on cell growth, heme content and δ-aminolevulinic acid dehydratase activity of malignant murine erythroleukemia cellsBiochemical and Biophysical Research Communications, 1979