SNO-hemoglobin is not essential for red blood cell-dependent hypoxic vasodilation.
The coupling of hemoglobin sensing of physiological oxygen gradients to stimulation of nitric oxide (NO) bioactivity is an established principle of hypoxic blood flow. One mechanism proposed to explain this oxygen-sensing-NO bioactivity linkage postulates an essential role for the conserved Cys93 residue of the hemoglobin beta-chain (betaCys93) and, specifically, for S-nitrosation of betaCys93 to form S-nitrosohemoglobin (SNO-Hb). The SNO-Hb hypothesis, which conceptually links hemoglobin and NO biology, has been debated intensely in recent years. This debate has precluded a consensus on physiological mechanisms and on assessment of the potential role of SNO-Hb in pathology. Here we describe new mouse models that exclusively express either human wild-type hemoglobin or human hemoglobin in which the betaCys93 residue is replaced with alanine to assess the role of SNO-Hb in red blood cell-mediated hypoxic vasodilation. Substitution of this residue, precluding hemoglobin S-nitrosation, did not change total red blood cell S-nitrosothiol abundance but did shift S-nitrosothiol distribution to lower molecular weight species, consistent with the loss of SNO-Hb. Loss of betaCys93 resulted in no deficits in systemic or pulmonary hemodynamics under basal conditions and, notably, did not affect isolated red blood cell-dependent hypoxic vasodilation. These results demonstrate that SNO-Hb is not essential for the physiologic coupling of erythrocyte deoxygenation with increased NO bioactivity in vivo.
Isbell
T Scott
TS
Department of Pathology and Center for Free Radical Biology, University of Alabama at Birmingham, Birmingham, Alabama 35294, USA.
Sun
Chiao-Wang
CW
Wu
Li-Chen
LC
Teng
Xinjun
X
Vitturi
Dario A
DA
Branch
Billy G
BG
Kevil
Christopher G
CG
Peng
Ning
N
Wyss
J Michael
JM
Ambalavanan
Namasivayam
N
Schwiebert
Lisa
L
Ren
Jinxiang
J
Pawlik
Kevin M
KM
Renfrow
Matthew B
MB
Patel
Rakesh P
RP
Townes
Tim M
TM
D000818:Animals; D004912:Erythrocytes / Q000473:pathology / Q000502:physiology*; D006439:Hemodynamics; D006454:Hemoglobins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D006801:Humans; D000860:Hypoxia / Q000473:pathology / Q000503:physiopathology*; D051379:Mice; D009566:Nitrates / Q000032:analysis / Q000378:metabolism; D009569:Nitric Oxide / Q000378:metabolism; D009573:Nitrites / Q000032:analysis / Q000378:metabolism; D015538:Nitrosation; D010100:Oxygen / Q000378:metabolism; D026403:S-Nitrosothiols / Q000032:analysis / Q000378:metabolism; D014664:Vasodilation / Q000502:physiology*
D006454:Hemoglobins; D009566:Nitrates; D009573:Nitrites; D026403:S-Nitrosothiols; C103574:S-nitrosohemoglobin; D009569:Nitric Oxide; D010100:Oxygen