Hemoglobin dissociation

Sigmoidal E ax Model. The sigmoidal E ax model, originally derived based on oxygen-hemoglobin dissociation kinetics,is a generalization of the Emax model. It describes the response vs. drug concentration... 

The four binding sites of hemoglobin exhibit positive coopera-tivity in binding O, leading to the sigmoid shape of hemoglobin dissociation curve. 

The oxygen contents and partial pressures are related by the hemoglobin dissociation curves for maternal and fetal bloods. We used a modified Hill equation (17, 18) ... 

Reactivity. Hemoglobin can exist ia either of two stmctural coaformatioas, corresponding to the oxy (R, relaxed) or deoxy (T, tense) states. The key differences between these two stmctures are that the constrained T state has a much lower oxygen affinity than the R state and the T state has a lower tendency to dissociate into subunits that can be filtered in the kidneys. Therefore, stabilization of the T conformation would be expected to solve both the oxygen affinity and renal excretion problems. 

In 1982 a study of the usefulness of DBBF in the production of a blood substitute was reported (99). A single modification achieved the dual goals of reduced oxygen affinity and restricted tetramer—dimer dissociation. This work was confirmed in 1987 (98). The product, called aa-hemoglobin, was formulated in Ringer s lactate. P q under physiologic conditions is 3.7 kPa (28.0 torr). Hill s parameter is 2.2, and the Bohr effect was reduced (100). Plasma retention was increased, and the product appeared to be less heterogeneous than some of the other derivatives under study. Its production was scaled up by Baxter Healthcare Corp., under contract to the U.S. Army. 

THE OXYGEN DISSOCIATION CURVES FOR MYOGLOBIN HEMOGLOBIN SUIT THEIR PHYSIOLOGIC ROLES... 

Hemoglobin carbamates account for about 15% of the CO2 in venous blood. Much of the remaining COj is carried as bicarbonate, which is formed in erythrocytes by the hydration of COj to carbonic acid (H2CO3), a process catalyzed by carbonic anhydrase. At the pH of venous blood, HjCOj dissociates into bicarbonate and a proton. 

Figure 6-9. The Bohr effect. Carbon dioxide generated in peripheral tissues combines with water to form carbonic acid, which dissociates into protons and bicarbonate ions. Deoxyhemoglobin acts as a buffer by binding protons and delivering them to the lungs. In the lungs, the uptake of oxygen by hemoglobin releases protons that combine with bicarbonate ion, forming carbonic acid, which when dehydrated by carbonic anhydrase becomes carbon dioxide, which then is exhaled.

An Increased dissociation of the hemoglobin molecule to monomers can be demonstrated after treatment with p-chloromer-curlbenzoate often, the unstable subunit will precipitate Irreversibly. 

The presence of Individual chains In a hemoglobin variant can also be demonstrated by electrophoresis at alkaline pH after the protein has been dissociated Into Its subunits through exposure to 6 M urea In the presence of 3-mercaptoethanol. The buffer is either a barbital buffer or a tris-EDTA-boric acid buffer, pH 8.0 - 8.6, and contains 6 M urea and 3-niercapto-ethanol. Dissociation of the hemoglobin Into subunits Is best accomplished In a mixture of 1 ml 10 g% Hb (or whole hemolysate), 4 ml 6 M urea barbital or tris-EDTA-boric acid buffer, and 1 to 1.5 ml 3-mercaptoethanol. After 30 minutes to 1 hour the sample Is subjected to cellulose acetate or starch gel electrophoresis. Each chain has a specific mobility and an alteration In electrophoretic mobility easily Identifies the abnormal chain. 

Measurement of free t-PA in plasma presents challenges in terms of preventing t-PA from complexing to PAI-1 released from platelets after blood collection. To dissociate any preformed t-PA-PAI-1 complex, the anticoagulant pH has to be close to 3.0. Even if blood is collected with an acidic anticoagulant, the blood pH will rise because of the powerful buffering action of hemoglobin. Thus, the pH of plasma has to be adjusted to 3.0 in order to dissociate the t-PA-PAI-I complex (115). 

The structures of the oxime complex Co(NO)(dmg)2 (84) and its diphenylglyoxime analog exhibit strongly bent Co—NO groups, which are easily oxidized to the corresponding nitro compound.353 A mechanistic study of NO transfer from Co(NO)(dmg)2 to hemoglobin established that the reaction involves NO association with the protein subsequent to dissociation of NO from the Co complex.354 This mechanism is also consistent with the observation of nitrato complexes in reactions... 

The P02 of blood is the major factor determining the amount of oxygen chemically combined with hemoglobin, or the percent of hemoglobin saturation. The relationship between these two variables is illustrated graphically by the oxyhemoglobin dissociation curve (see Figure 17.6). This relationship is not... 

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