The red blood cell proteome and interactome identify a Band 3-BLVRB axis regulating hypoxic metabolic adaptation
Abstract
Red blood cells (RBCs) are transcriptionally silent yet dynamically remodel metabolism in response to oxygen tension. Using ultra-pure human RBCs, we generated the deepest contamination-free proteome to date (3,775 proteins) and mapped the oxygen-dependent interactome. These datasets reveal an oxygen-responsive metabolon centered on the Band 3 (SLC4A1) N-terminus. We identify biliverdin reductase B (BLVRB) as a previously unrecognized Band 3 interactor that dissociates under hypoxia, coincident with increased Band 3-deoxyhemoglobin contacts. This reversible assembly functions as an oxygen-sensitive switch coordinating redox and glycolytic remodeling. Humanized mice lacking Band 3 N-terminal segments exhibit impaired oxygen-dependent regulation of BLVRB binding to band 3, impaired hypoxic activation of glycolysis, reduced 2,3-bisphosphoglycerate synthesis, and diminished exercise tolerance, demonstrating physiological relevance. Population-scale cis-pQTLs for SLC4A1 and BLVRB suggest functions beyond canonical heme catabolism. Mechanistically, biochemical analyses in vitro suggest that hemoglobin β (HBB), Band 3, and BLVRB can undergo S-nitrosation and may participate in trans-nitrosation reactions with the glycolytic enzyme GAPDH, whose modification at C152 inhibits enzymatic activity in vitro. Collectively, these findings define a Band 3-BLVRB axis that integrates oxygen-dependent protein interactions with thiol-based redox chemistry, providing a framework for understanding how an anucleate cell achieves metabolic adaptability through reversible protein-protein interactions and post-translational modification. These findings suggest that perturbation of the Band 3-BLVRB axis may influence oxygen delivery and metabolic flexibility during hypoxic stress, with potential relevance to high-altitude adaptation, exercise physiology, and cardiopulmonary disease.
Article Details
Authors (29)
Aaron Vahe Issaian
University of Colorado Anschutz Medical Campus, Aurora, Colorado, United States
Monika Dzieciatkowska
Shaun Bevers
University of Colorado - Anschutz Medical Campus, Aurora, Colorado, United States
Zohreh Safari
University of Maryland School of Medicine, Baltimore, Maryland, United States
Ariel M Hay
University of Virginia, Charlottesville, Virginia, United States
Francesca I Cendali
University of Colorado Anschutz Medical Campus, Aurora, Colorado, United States
Amy Argabright
University of Colorado Anschutz Medical Campus, Aurora, CO, USA, Aurora, Colorado, United States
Stephen C Rogers
University of Maryland School of Medicine, Baltimore, Maryland, United States
Anthony J Saviola
University of Colorado Anschutz Medical Campus, Aurora, Colorado, United States
Jasmina S Redzic
University of Colorado Denver, Aurora, Colorado, United States
Eric Wartchow
Children's Hospital Colorado, Aurora, Colorado, United States
Julie Reisz Haines
University of Colorado, Anschutz Medical Center, Aurora, Colorado, United States
Gregory R Keele
RTI International, Research Triangle Park, North Carolina, United States
Zachary B Haiman
University of Colorado Denver - Anschutz Medical Campus, Denver, Colorado, United States
Travis Nemkov
Daniel Stephenson
Christina Lisk
University of Colorado Anschutz Medical Campus, Aurora, Colorado, United States
Francesca Vallese
Bernhard O. Palsson
University of California-San Diego School of Medicine, La Jolla, California, United States
S Bruce King
Wake Forest University, Winston-Salem, North Carolina, United States
Grier P Page
RTI International, Atlanta, Georgia, United States
Allan Doctor
University of Maryland School of Medicine, Baltimore, Maryland, United States
Krystalyn E Hudson
Columbia University, New York, New York, United States
Kirk C Hansen
University of Colorado Anschutz Medical Campus School of Medicine, Aurora, Colorado, United States
David C. Irwin
University of Colorado Health Science Center, Aurora, Colorado, United States
Narla Mohandas
James C Zimring
Canadian Blood Services, Ottawa, Ontario, Canada
Elan Z Eisenmesser
University of Colorado - Anschutz Medical Campus, Colorado, Colorado, United States
Angelo D'Alessandro