Nitric oxide-mediated heme oxidation and selective beta-globin nitrosation of hemoglobin from normal and sickle erythrocytes.
Nitric oxide (NO) has been reported to modulate the oxygen affinity of blood from sickle cell patients (SS), but not that of normal adult blood (AA), with little or no heme oxidation. However, we had found that the NO donor compounds 2-(N, N-diethylamino)-diazenolate-2-oxide (DEANO) and S-nitrosocysteine (CysNO) caused increased oxygen affinity of red cells from both AA and SS individuals and also caused significant methemoglobin (metHb) formation. Rapid kinetic experiments in which HbA(0), AA, or SS erythrocytes were mixed with CysNO or DEANO showed biphasic time courses indicative of initial heme oxidation followed by reductive heme nitrosylation, respectively. Hemolysates treated with CysNO showed by electrospray mass spectrometry a peak corresponding to a 29 mass unit increase (consistent with NO binding) of both the beta(A) and beta(S) chains but not of the alpha chains. Therapeutic use of NO in sickle cell disease may ultimately require further optimization of these competing reactions, i.e., heme reactivity (nitrosylation or oxidation) versus direct S-nitrosation of hemoglobin on the beta-globin.
Hrinczenko
B W
BW
Laboratory of Chemical Biology and Structural Mass Spectroscopy Group, National Institutes of Diabetes and Digestive and Kidney Diseases, Bethesda, Maryland 20892, USA.
Schechter
A N
AN
Wojtkowski
T L
TL
Pannell
L K
LK
Cashon
R E
RE
Alayash
A I
AI
D000755:Anemia, Sickle Cell / Q000378:metabolism*; D003545:Cysteine / Q000031:analogs & derivatives / Q000378:metabolism; D004048:Diethylamines / Q000378:metabolism; D004912:Erythrocytes / Q000378:metabolism* / Q000473:pathology; D005914:Globins / Q000378:metabolism*; D006418:Heme / Q000378:metabolism*; D006441:Hemoglobin A / Q000378:metabolism; D006451:Hemoglobin, Sickle / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D013058:Mass Spectrometry; D008706:Methemoglobin / Q000378:metabolism; D009569:Nitric Oxide / Q000378:metabolism*; D009589:Nitrogen Oxides; D009603:Nitroso Compounds / Q000378:metabolism; D010084:Oxidation-Reduction; D010100:Oxygen / Q000378:metabolism; D026403:S-Nitrosothiols; D013053:Spectrophotometry; D013816:Thermodynamics
D004048:Diethylamines; D006451:Hemoglobin, Sickle; D009589:Nitrogen Oxides; D009603:Nitroso Compounds; D026403:S-Nitrosothiols; D009569:Nitric Oxide; D006418:Heme; D005914:Globins; D008706:Methemoglobin; D006441:Hemoglobin A; C021315:S-nitrosocysteine; C087229:diethylamine dinitric oxide adduct; D003545:Cysteine; D010100:Oxygen