Autoxidation of extracellular oxyhaemoglobin from the polychaete Perinereis brevicirris (Grube)

doi:10.1016/0305-0491(83)90033-0

Comparative Biochemistry and Physiology Part B: Comparative Biochemistry

Volume 75, Issue 1, 1983, Pages 17-21

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Abstract

1.
1. SDS-polyacrylamide gel electrophoresis showed that the Perineis haemoglobin consisted of three subunits with mol. wt of 12,000 (subunit 1), 40,000 (subunit 2) and 54,000 (subunit 3) in a molar ratio of 1:4:3.
2.
2. At alkaline pH, the oxyhaemoglobin dissociates into subunits. Therefore, the two haem-containing, subunits 1 and 2, could be separated by gel filtration on a Sephadex G-75 column.
3.
3. The autoxidation rates of oxyhaemoglobin and the oxy-forms of the haem-containing subunits to met-forms were measured in the pH range of 5.4–11.6 and the pH dependencies of their autoxidation rates were compared with that of vertebrate oxymyoglobin.
4.
4. The autoxidation rates of subunits 1 and 2 were approx 500 and 30 times, respectively, that of the intact haemoglobin at pH 7.5.
5.
5. The activation energy of the autoxidation reaction of the whole oxyhaemoglobin was calculated to be 30.5 kcal/mol at pH 7.2.

References (21)

W.D. Brown et al.
Autoxidation of oxymyoglobins
J. biol. Chem.
(1969)
E. Chiancone et al.
Physicochemical and functional properties of Perinereis cultrifera (Grube) erythrocruorin
Biochim. biophys. Acta
(1977)
R.L. Garlick et al.
The amino acid sequence of a major polypeptide chain of earthworm haemoglobin
J. biol. Chem.
(1982)
T. Gotoh et al.
Autoxidation of native oxymyoglobin from bovine heart muscle
Archs Biochem. Biophys.
(1974)
Y. Satoh et al.
Autoxidation of oxymyoglobin
J. biol. Chem.
(1981)
K. Shikama et al.
Aplysia myoglobin with an unusual haem environment
Biochim. biophy. Acta
(1982)
T. Suzuki et al.
Amino acid sequence of the smallest polypeptide chain containing haem of extracellular haemoglobin from the polychaete Tylorrhynchus heterochaetus
Biochim. biophys. Acta
(1982)
S.N. Vinogradov et al.
The dissociation of annelid extracellular haemoglobins and their quaternary structure
Comp. Biochem. Physiol.
(1980)
W.J. Wallace et al.
The mechanisms of haemoglobin autoxidation
K. Weber et al.
The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis
J. biol. Chem.
(1969)

There are more references available in the full text version of this article.

Cited by (8)

Autoxidation studies of extracellular hemoglobin of Glossoscolex paulistus at pH 9: Cyanide and hydroxyl effect
2005, Biophysical Chemistry
The complex oligomeric assembly of the hemoglobin subunits may influence the autoxidation rate. To understand this relation, the rate of autoxidation was studied at pH 9.0, where the Glossoscolex paulistus Hemoglobin (GpHb) dissociates. At alkaline pH, this hemoglobin is dissociated into monomers, trimers and tetramers, allowing the study of the integral protein and monomer subunit autoxidation on independent experiments. The autoxidation rate was evaluated in the presence and absence of cyanide (CN⁻), a strong field ligand to the ferric ion. The oxidation kinetic was monitored using the UV–vis absorption at 415 nm, and resulted in: i) bi-exponential kinetics for the whole hemoglobin (indicating a fast and a slow oxidative process) and ii) mono-exponential for the monomer (indicating a single process). To understand the specific characteristics of each autoxidation process, Arrhenius plots allowed the determination of the activation energy. The experimental results indicate for the whole hemoglobin in the absence of CN⁻ an activation energy of 150±10 kJ mol⁻¹ for the fast and the slow processes. Under the same conditions the monomer displayed an activation energy of 160±10 kJ mol⁻¹, very close to the value obtained for the integral protein. The pseudo-second order rate constant for the whole protein autoxidation by CN⁻ showed two different behaviors characterized by a rate constant k_CN1′=0.11±0.02 s⁻¹ mol⁻¹ L for CN⁻ concentrations lower than 0.012 mol L⁻¹; and k_CN1ʺ=0.76±0.04 s⁻¹ mol⁻¹ L at higher concentrations for the fast process, while the slow process remain constant with k_CN2=0.033±0.002 s⁻¹ mol⁻¹ L. The monomer has a characteristic rate constant of 0.041±0.002 s⁻¹ mol⁻¹ L for all cyanide concentrations. Comparing the results for the slow process of the whole hemoglobin and the oxidation of the monomer, it is possible to infer that the slow process has a strong contribution of the monomer in the whole hemoglobin kinetic. Moreover, as disulfide linkers sustain the trimer assembly, cooperativity may explain the higher kinetic constant for this subunit.
Unusual pattern of haemoglobin tissue specificity in the "red maggot" worm Hyboscolex longiseta (polychaeta, scalibregmidae)
1988, Comparative Biochemistry and Physiology -- Part B: Biochemistry and
- 1.
  1. Hyboscolex longiseta has a typical Annelid extracellular haemoglobin present in blood lacunae surrounding the gut, but no external gills. In EDTA-containing buffers this vascular haemoglobin dissociates into 133,000–139,000 components which correspond to 1/24th's of the original high mol. wt haemoglobin and which dissociate further when treated with 2-mercaptoethanol.
- 2.
  2. Hyboscolex longiseta has a minor dimeric “myoglobin” and a major tetrameric “myoglobin”, the letter being identical to the haemoglobin contained in coelomic erythrocytes. There is also in muscle extracts a trace of what may be the original basic true myoglobin.
- 3.
  3. Haemoglobin in muscle extracts has a low oxygen affinity, whereas haemoglobin in the vascular system has a high oxygen affinity. Thus, H. longiseta has evolved a reverse oxygen transfer system by a combination of increasing the oxygen affinity of the vascular haemoglobin and by changing the site of tissue expression of the erythrocyte haemoglobin so that much more of that protein is actually made in the body wall musculature as a “myoglobin”.
- 4.
  4. High oxygen affinity respiratory proteins surrounding the gut are postulated to function in keeping low oxygen partial pressures. In this way polychaetes (which are part of the decomposer food chain) utilise detritus and associated microorganisms. Bacteria produce alcohols and other compounds, both from aerobic breakdown of lignins and other recalcitrant molecules and from anaerobic metabolism.
Auto-deoxygenation of annelid giant haemoglobin in vitro
1984, International Journal of Biological Macromolecules
Mammalian tetrameric oxyhaemoglobin does not deoxygenize under air-saturated conditions. But, the absorption spectrum of purified giant oxyhaemoglobin from the polychaete Perinereis brevicirris changed from the oxy form to the deoxy form when incubated in 50 mM buffer (pH 7) for 15h at 26°C under air-saturated conditions. This unusual reaction, namely autodeoxygenation, is characterized by the sigmoidal time course of the reaction, involving an initial slow reaction accompanied by a subsequent rapid reaction, and the formation of turbidity of the solution during the reaction. In this communication, we reprot for the first time the effects of Ca²⁺, Mg²⁺, EDTA, haemoglobin concentration and temperature on the rate of auto-deoxygenation.
Separation of constituent polypeptide chains containing heme from extracellular hemoglobin of the polychaete Perinereis brevicirris (Grube)_
1983, Comparative Biochemistry and Physiology -- Part B: Biochemistry and
- 1.
  1 Perinereis cyanomethemoglobin was reduced with 1 mM DTT and applied to a column of Sephadex G-100 equilibrated with 5 mM phosphate buffer (pH 7.2) containing 1 mM DTT. The polypeptides eluted in the position corresponding to a mol. wt of about 18,000. Each retained heme without any appreciable loss.
- 2.
  2. These polypeptides were separated into six fractions on a column of DEAE-Sepharose CL-6B equilibrated with 5 mM phosphate buffer (pH 7.2) containing 1 mM DTT. Each polypeptide showed a clear absorbance maximum in the Soret region.
- 3.
  3. The amino acid compositions and NH₂-terminal sequences of these polypeptides were determined.
- 4.
  4. The procedure developed for separation of constituent polypeptides with attached heme was applicable to other annelid extracellular hemoglobins; i.e. those of Tylorrhynchus and Neanthes.
- 5.
  5. In contrast to previous suggestions based on heme analysis, these results strongly suggest that each constituent polypeptide chain of polychaete hemoglobins contains heme.
Hemoglobin Extracted From Perinereis aibuhitensis Acts as a Cell Culture Medium Supplement to Reduce Apoptosis
2021, Frontiers in Marine Science
A comparative study on earthworm hemoglobins: An amino acid sequence comparison of monomer globin chains of two species, Pontodrilus matsushimensis and Pheretima communissima that belong to the family megascolecidae
1996, Zoological Science

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Comparative Biochemistry and Physiology Part B: Comparative Biochemistry

Abstract

References (21)

Autoxidation of oxymyoglobins

J. biol. Chem.

Physicochemical and functional properties of Perinereis cultrifera (Grube) erythrocruorin

Biochim. biophys. Acta

The amino acid sequence of a major polypeptide chain of earthworm haemoglobin

J. biol. Chem.

Autoxidation of native oxymyoglobin from bovine heart muscle

Archs Biochem. Biophys.

Autoxidation of oxymyoglobin

J. biol. Chem.

Aplysia myoglobin with an unusual haem environment

Biochim. biophy. Acta

Amino acid sequence of the smallest polypeptide chain containing haem of extracellular haemoglobin from the polychaete Tylorrhynchus heterochaetus

Biochim. biophys. Acta

The dissociation of annelid extracellular haemoglobins and their quaternary structure

Comp. Biochem. Physiol.

The mechanisms of haemoglobin autoxidation

The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis

J. biol. Chem.

Cited by (8)

Autoxidation studies of extracellular hemoglobin of Glossoscolex paulistus at pH 9: Cyanide and hydroxyl effect

Unusual pattern of haemoglobin tissue specificity in the "red maggot" worm Hyboscolex longiseta (polychaeta, scalibregmidae)

Auto-deoxygenation of annelid giant haemoglobin in vitro

Separation of constituent polypeptide chains containing heme from extracellular hemoglobin of the polychaete Perinereis brevicirris (Grube)_

Hemoglobin Extracted From Perinereis aibuhitensis Acts as a Cell Culture Medium Supplement to Reduce Apoptosis

A comparative study on earthworm hemoglobins: An amino acid sequence comparison of monomer globin chains of two species, Pontodrilus matsushimensis and Pheretima communissima that belong to the family megascolecidae