Wake Forest Physics
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WFU Physics Colloquium
TITLE: Effects of Red Blood Cells on Nitric Oxide Bioactivity
SPEAKER: Xiaohua Liu
TIME: Monday April 24, 2017 at 10:00 AM
PLACE: ZSR 204
ABSTRACT
Red blood cells (RBCs) as one of important cells in human blood vessel
are abundant in hemoglobin (Hb). Hb has four heme ferrous sites for
binding oxygen, nitric oxide (NO), carbon monoxide (CO) and other small
molecules. NO as an endothelial derived relaxing factor is involved in
many physiological and pathological processes. Its reaction with Hb is
extremely fast. Because of RBCs’ unique biconcave shape and intrinsic
membrane barrier, the rate of NO scavenging by encapsulated Hb in RBCs
is limited and slower 1000 times than cell-free hemoglobin. During blood
storage, the shape of RBCs tends to spherical conversion and membrane
deformability reduced. Hemolysis from RBCs breaking down leads to low NO
bioavailability. It is supposed that old stored RBCs scavenging more NO
than young ones. We designed competition experiments and stop-flow
spectroscopy to determine that older stored RBCs scavenging NO 1.7-1.8
times faster than fresh stored RBCs. Through computational simulation,
to obtain this observed result, the RBC membrane permeability to NO has
to increase 5 to 70 folds. Under hypoxic and acidic condition, nitrite
is reduced to NO by deoxygenated hemoglobin. Since two states of Hb
(R-state and T-state) are reversible under proper condition, Hb acts as
an allosterically controlled nitrite reductase. Using electron
paramagnetic resonance spectroscopy, chemiluminescent detection and
platelet activation study, we confirmed that Hb plays primary role for
nitrite reduction in human body. Meanwhile, nutrients also can modulate
NO export. The nitrite bioactivation in RBCs senses to low oxygen and
low nutrients areas. Due to the rate constant of Hb and NO reaction, NO
formed in the RBC is not likely to escape scavenging. NO activity is
preserved in as a nitrosothiol through a nitrosylation process. Our
result found that the nitrite bioactivation is involved surface protein
nitrosation. Nitric oxide synthase (NOS) is an enzyme for converting
L-arginine to NO and citrulline. Arginase as another enzyme converts
L-arginine to urea and L-ornithine. Nor-NOHA as arginase inhibitor is
supposed to block the arginase pathway so that increase NO production.
However, our data suggest that nor-NOHA reacts with hemoglobin to form
nitrite and nitrate not through NO production.
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