Dissociation of the extracellular hemoglobin of Arenicola cristata

Vinogradov, Serge N.; Shlom, Jerald M.; Doyle, M. A.

doi:10.1016/0305-0491(80)90123-6

Cited by 6 publications

(3 citation statements)

References 16 publications

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“…However, LtEc should not be stored at higher pH levels, since its thermal stability significantly decreases at alkaline pH (T m ¼ 45 C at pH 8.0). This effect has also been observed with other Ec's, including Glossoscolex paulistus, Planorbis corneus, and Arenicola cristata (Bispo et al 2005, Vinogradov et al 1980, Wood and Mosby 1975.…”

Section: Discussionsupporting

confidence: 70%

Prolonging the shelf life ofLumbricus terrestriserythrocruorin for use as a novel blood substitute

Muzzelo

Neely

Shah

et al. 2017

Artificial Cells, Nanomedicine, and Biotechnology

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Limitations associated with the storage of red blood cells have motivated the development of novel blood substitutes that are able to withstand long-term storage at elevated temperatures. The hemoglobin of the earthworm Lumbricus terrestris (LtEc) is an attractive blood substitute candidate, since it is resistant to oxidation and aggregation during storage. Several factors were investigated to optimize the thermal and oxidative stability of LtEc during storage, including pH, antioxidant supplements, and deoxygenation. A strategy for the reduction of fully oxidized LtEc with antioxidants was also developed. Overall, LtEc was shown to have the highest thermal stability in Ringer's Modified Lactate solution with 10 mM HEPES at pH 7.0. Deoxygenation of the LtEc was also shown to significantly reduce oxidation of the ferrous heme iron (e.g., %Feafter 7 d at 37 °C = 75.7%). However, even in cases where oxidation does occur, the addition of 1.8 mM ascorbic acid (AA) was found to reduce 98.3% of the oxidized LtEc (37 μM heme). Most importantly, the oxygen transport properties of LtEc were unaffected by storage at high temperatures or oxidation followed by reduction with AA. These results show that LtEc can be stored at high temperatures (37 °C) without any significant loss of function.

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Section: Discussionsupporting

confidence: 70%

Prolonging the shelf life ofLumbricus terrestriserythrocruorin for use as a novel blood substitute

Muzzelo

Neely

Shah

et al. 2017

Artificial Cells, Nanomedicine, and Biotechnology

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“…The resistance of intrasubunit disulfide bonds to reducing agents could explain the underestimation of subunit mass by SDS/PAGE. Compared to fully reduced proteins, unreduced proteins containing intrachain disulfide bonds possess a more compact hydrodynamic shape in SDS and therefore exhibit a higher than expected electrophoretic mobility [35,36].…”

Section: Resultsmentioning

confidence: 99%

Native and subunit molecular mass and quarternary structure of the hemoglobin from the primitive branchiopod crustacean Triops cancriformis

et al. 2006

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The physicochemical features of the large-sized, extracellular hemoglobins (Hbs) of branchiopod crustaceans have engaged structural biologists and animal physiologists since Svedberg's sedimentation analysis of the Many branchiopod crustaceans are endowed with extracellular, highmolecular-weight hemoglobins whose exact structural characteristics have remained a matter of conjecture. By using a broad spectrum of techniques, we provide precise and coherent information on the hemoglobin of one of the phylogenetically 'oldest' extant branchiopods, the tadpole shrimp Triops cancriformis. The hemoglobin dissociated under reducing conditions into two subunits, designated TcHbA and TcHbB, with masses of 35 775 ± 4 and 36 055 ± 4 Da, respectively, determined by ESI-MS. Nonreducing conditions showed only two disulfide-bridged dimers, a homodimer of TcHbA, designated D1 (71 548 ± 5 Da), and the heterodimer D2 (71 828 ± 5 Da). Carbamidomethylation of free SH groups revealed the presence of three cysteines per subunit and indicated one intrasubunit and one intersubunit disulfide bridge. Ultracentrifugation and light-scattering experiments under nondenaturating conditions yielded mass estimates that suggested an uneven number of 17 subunits forming the native hemoglobin. This unrealistic number resulted from the presence of two size classes (16-mer and 18-mer), which were recognized by native PAGE and Ferguson plot analysis. ESI-MS revealed three hemoglobin isoforms with masses of 588.1 kDa, 662.0 kDa, and 665.0 kDa. The 16-mer and the smaller 18-mer species are supposed to be composed of TcHbA only, given the dominance of this subunit type in SDS ⁄ PAGE. Transmission electron microscopy of negatively stained specimens showed a population of compact molecules with geometrical extensions of 14, 16 and 9 nm. The proposed stoichiometric model of quarternary structure provides the missing link to achieve a mechanistic understanding of the structure-function relationships among the multimeric arthropodan hemoglobins.Abbreviations CI, confidence interval; D 20 , diffusion coefficient for 20°C; DLS, dynamic light scattering; Hb, hemoglobin; MALLS, multiangle laser-light scattering; s 20,w , sedimentation coefficient for 20°C in water; TEM, transmission electron microscopy; m, partial specific volume.

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“…Protons are also important effectors in extracellular Hbs and have been studied by several groups. Although various reports have described the dissociation of oligochaete Hb at alkaline pH [26,27], including L. terrestris [28,29], Eisenia foetida [30,31], and two species of Pheretima [32], and of polychaetes such as Arenicola [33,34] and Abarenicola affinis affinis [35], very little is known about the process of association in these proteins and the role of protons. The main objective of this study was therefore to examine the thermodynamic features of the pressure dissociation of extracellular Hb at alkaline pH, with particular emphasis on quantification of the protons involved in the dissociation.…”

Section: Introductionmentioning

confidence: 99%

pH dependence of the dissociation of multimeric hemoglobin probed by high hydrostatic pressure

Bispo

Santos

Landini

et al. 2007

Biophysical Chemistry

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Dissociation of the extracellular hemoglobin of Arenicola cristata

Cited by 6 publications

References 16 publications

Prolonging the shelf life ofLumbricus terrestriserythrocruorin for use as a novel blood substitute

Prolonging the shelf life ofLumbricus terrestriserythrocruorin for use as a novel blood substitute

Native and subunit molecular mass and quarternary structure of the hemoglobin from the primitive branchiopod crustacean Triops cancriformis

pH dependence of the dissociation of multimeric hemoglobin probed by high hydrostatic pressure

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