Deer Hemoglobin Research Articles

Entropy-Driven Intermediate Steps of Oxygenation May Regulate the Allosteric Behavior of Hemoglobin

When the oxygen binding isotherms of human, bovine and fallow deer (Dama-Dama) hemoglobins are measured at different temperatures either by optical or calorimetric techniques, analyses according to the Adair’s formalism show that at least one of the intermediate steps of ligation has a positive enthalpy change, i.e., absorbs rather than emitting heat, indicating that it is entropy rather than enthalpy driven (Bucci, E., et al. 1991. Biochemistry. 30:3195–3199; Bucci, E., et al. 1993. Biochemistry. 32:3519–3526; Johnson, C., et al. 1992. Biochemistry. 31:10074–10082; Johnson, C., et al. 1995. Biophys. Chem. 59:107–117). This phenomenon is confirmed in systems in which the β82 lysines of human hemoglobin are covalently cross-linked by acylation with dicarboxylic acids of increasing length, namely the fumaryl (four-carbon-long), adipoyl (six-carbon-long), and sebacoyl (10-carbon-long) residues. Consistently in all of the systems here reported, the enthalpy excursions are masked by compensatory entropy changes, which keep the free energy of ligand binding constant for the first three steps of oxygenation. Furthermore, the cooperativity index and the overall oxygen affinity seem to be correlated to the positive enthalpy excursions of the intermediate steps of ligation. Fumaryl-Hb (hemoglobin cross-linked with a fumaryl residue, four carbons) with the lowest absorption of heat has the highest affinity and lowest cooperativity index. Adipoyl-Hb (hemoglobin cross-linked with an adipoyl residue, six carbons) has the highest absorption of heat and the highest cooperativity index. It appears that nonuniform heat release by the intermediates of oxygenation is part of the allosteric phenomena in hemoglobin systems. There is not enough information that would allow assigning these phenomena to the interplay of the various conformations described for hemoglobin besides the classic T (Fermi et al. 1984. J. Mol. Biol. 175:159–174) and R (Shanaan. 1983. J. Mol. Biol. 171:31–59), as listed at the end of the Discussion. The possibility cannot be excluded that entropy-driven steps characterize new conformational transitions still to be described.

Ultrasonic absorption in deer blood

The erythrocyte sickling in white‐tailed deer is very similar to that in humans. Deer hemoglobin molecules polymerize at high pH and high oxygen tension. Ultrasonic absorption in deer blood versus pH and oxygen tension has been measured with Carstensen's fix path method and a method described in an earlier paper [K. K. Shung et al., Blood 54, 451–458 (1979)]. It has been found that the ultrasonic specific absorption at 15 MHz increases from 27 dB‐cc/ g‐cm to 66 dB‐cc/g‐cm when the pH of the blood is increased from 7 to 8 at an oxygen saturation of 95%. This result compares favorably with the data for humans which show that, under physiological conditions favoring sickling, the specific absorption at 15 MHz is around 60 dB‐cc/g‐cm. It was also found that ultrasonic absorption starts to increase at oxygen saturation around 50% at high pH and does not change appreciably at physiological pH. [Work supported by NSF grant DAR‐8018415 and NIH grant HL26871.]

Crystallography and oriented single crystal electron microscopy of hemoglobin deer II, a hemoglobin that exhibits matchstick-shaped erythrocytes

Deer hemoglobin beta chain type II has been crystallized and preliminary diffraction data and oriented single crystal transmission electron micrographs have been obtained. The crystals are monoclinic P2 1 with Z = 4. The electron micrographs show a herringbonelike structure in the ab plane with open rectangular solvent channels and a fiber-like arrangement of molecules perpendicular to this plane.

Application of a restrained least-squares refinement procedure to sickling deer hemoglobin

Application of a restrained least-squares refinement procedure to sickling deer hemoglobin

Viscosity and gelation studies in deer hunting hemoglobins.

Sickling, viscosity and gelling properties of the red cells and the hemoglobins of three Virginia white-tailed deer homozygous for types II and III (the sickling types) and V (the nonsickling type), respectively, have been analyzed. The sickling of erythrocytes of deer with type II or III is inhibited by urea and cyanate at concentrations which are comparable to those used in in vitro studies of red cells from patients with sickle cell anemia. No differences were observed between the viscosities of the three deer hemoglobin types at temperatures of 12 degrees C or above. High concentrations of deer hemoglobin types II and III gelled at 1 degree C and at pH values of 7.4-7.7; the minimum gelling concentration of type II was 33.5 g% and of type III was 38 g%. Gel formation was not observed at pH values between 6.7-7.1. Hemoglobin type V did not gel and prevented the formation of gels of type II and III in mixtures at pH 7.6-7.7.

Oxygen transport and phosphorylated glycolytic intermediates of the ruminant blood—I. sika deer ( Cervus nippon nippon)

1. 1. Values for several hematologic parameters were determined for sika deer ( Cervus nippon nippon), the most prominent features of which were a high red cell count, (10·56±2·64) × 10 6/μl, and a low MCV, 39·3 ± 6· μ 3. 2. 2. The hemoglobin of sika deer was a single component of the same mobility when examined by starch gel electrophoresis. 3. 3. The oxygen affinity of deer blood was slightly lower than that of human adult blood through the physiological pH range (pH 6·9 ∼ 7·7). 4. 4. There was no significant difference in the intra- vs extra-cellular pH relationship between deer and human adult blood. 5. 5. Concentrations of 2,3-diphosphoglycerate (2,3-DPG) and the other organic phosphates were very low in deer red cells. 6. 6. It is concluded that the lower oxygen affinity of deer blood in the essential absence of co-factor phosphates such as 2,3-DPG might be of an intrinsic nature reflecting the characteristic primary structure of the deer hemoglobin itself.

Structural Comparison of Polymorphic Hemoglobins of Deer with Those of Sheep and Other Species

Abstract The structure of deer hemoglobin III, associated with sickling of the erythrocytes of Florida white tailed deer, is compared with that of deer hemoglobin V, the presence of which precludes sickling of erythrocytes under appropriate conditions, by peptide mapping and amino acid analysis techniques. Peptide mapping techniques, including combined specific color reactions, have been utilized to relate hemoglobin peptides in the deer hemoglobins to those of several species for which the amino acid sequence is known. Certain structural regions of the α and β polypeptide chains of deer hemoglobin are closely homologous to the corresponding regions in the α and β chains of these other species, as illustrated by a graph based upon the peptide mapping studies and known sequences. The tryptic peptide mapping pattern of the α chains of deer hemoglobins III and V is nearly identical with that of the sheep α chains from the three polymorphic sheep hemoglobins. In contrast to the similarity of the α chains of these two species, the β chains exhibited numerous inter- and intraspecies differences. However, the marked variations of the β chains among the polymorphic hemoglobins of the ruminants are largely confined to distinct regions of these β chains. Although the significance of a linear comparison of the homologous regions is not apparent, the relationship of these regions is clarified by use of a three-dimensional model.

Hemoglobin Polymorphism in White-Tailed Deer: Subunit Basis

Abstract A variety of aberrant erythrocyte forms have been related to seven adult and two fetal hemoglobins in white-tailed deer. While sickling of the erythrocyte was not associated with a single hemoglobin type, it was precluded by hemoglobin V or VII, even when in combination with other hemoglobin types normally associated with sickling. The subunit basis of the hemoglobin polymorphism was presented. Two kinds of α subunits, six kinds of β subunits and one γ subunit were related to the whole hemoglobin molecule. The heterogeneity of the deer hemoglobins was based upon a variety of combinations of these numerous polypeptide chains. It was concluded from the results of limited structural studies that there were multiple peptide differences upon comparison of three non-α polypeptide chains.

The Sickling Phenomenon and Heterogeneity of Deer Hemoglobin.

SummaryA marked degree of heterogeneity is demonstrable in deer hemoglobin. This heterogeneity appears to be on a genetic basis. A single hemoglobin component responsible for sickling of deer hemoglobin can be identified.Addendum. Since this paper was prepared, similar electrophoretic results were reported by Kitchen, H., Putnam, F. W. and Taylor, W. J., in Science, 1964, v144, 1237.

Tactoid formation in deer hemoglobin

Sickle cell preparations were made on eight Virginia white-tail deer ( Odocoileus virginianus) killed during the 1958–59 hunting season. All preparations showed sickling. Paper electrophoresis of carboxyhemoglobin from these animals revealed identical mobilities and a single hemoglobin component present at pH 8.6. Alkali denaturation of the deer carboxyhemoglobin solution showed that it was markedly resistant to alkali denaturation and that the kinetics of the denaturation were different from that of human cord hemoglobin. Deer carboxyhemoglobin migrated 0.6 x 10–5 cm2/volt/sec.–1 faster than human carboxyhemoglobin A in moving boundary electrophoresis in veronal buffer at pH 8.2. Tactoids were demonstrated in free hemoglobin solution with phase microscopy after concentration of the solution. These particles are very similar morphologically to these prepared from hemoglobin solution containing hemoglobin S.