Faculty Collaboration Database

Human red blood cells at work: identification and visualization of erythrocytic eNOS activity in health and disease. Blood 2012 Nov 15;120(20):4229-37

Date

09/26/2012

Pubmed ID

23007404

DOI

10.1182/blood-2012-07-442277

Scopus ID

2-s2.0-84869786258 (requires institutional sign-in at Scopus site)   142 Citations

Abstract

A nitric oxide synthase (NOS)-like activity has been demonstrated in human red blood cells (RBCs), but doubts about its functional significance, isoform identity and disease relevance remain. Using flow cytometry in combination with the nitric oxide (NO)-imaging probe DAF-FM we find that all blood cells form NO intracellularly, with a rank order of monocytes > neutrophils > lymphocytes > RBCs > platelets. The observation of a NO-related fluorescence within RBCs was unexpected given the abundance of the NO-scavenger oxyhemoglobin. Constitutive normoxic NO formation was abolished by NOS inhibition and intracellular NO scavenging, confirmed by laser-scanning microscopy and unequivocally validated by detection of the DAF-FM reaction product with NO using HPLC and LC-MS/MS. Using immunoprecipitation, ESI-MS/MS-based peptide sequencing and enzymatic assay we further demonstrate that human RBCs contain an endothelial NOS (eNOS) that converts L-(3)H-arginine to L-(3)H-citrulline in a Ca(2+)/calmodulin-dependent fashion. Moreover, in patients with coronary artery disease, red cell eNOS expression and activity are both lower than in age-matched healthy individuals and correlate with the degree of endothelial dysfunction. Thus, human RBCs constitutively produce NO under normoxic conditions via an active eNOS isoform, the activity of which is compromised in patients with coronary artery disease.

Author List

Cortese-Krott MM, Rodriguez-Mateos A, Sansone R, Kuhnle GG, Thasian-Sivarajah S, Krenz T, Horn P, Krisp C, Wolters D, Heiß C, Kröncke KD, Hogg N, Feelisch M, Kelm M

Author

Neil Hogg PhD Associate Dean, Professor in the Biophysics department at Medical College of Wisconsin

MESH terms used to index this publication - Major topics in bold

Adult
Amino Acid Sequence
Arginine
Chromatography, High Pressure Liquid
Citrulline
Coronary Artery Disease
Endothelium, Vascular
Erythrocytes
Flow Cytometry
Fluoresceins
Fluorescent Dyes
Humans
Immunoprecipitation
Mass Spectrometry
Microscopy, Confocal
Molecular Sequence Data
NG-Nitroarginine Methyl Ester
Nitric Oxide
Nitric Oxide Synthase Type III
Oxyhemoglobins
Sequence Alignment
Sequence Analysis, Protein
Sequence Homology, Amino Acid