Small-angle X-ray studies of Lumbricus terrestris haemoglobin

Pilz, Ingrid; Schwarz, Erika; Vinogradov, Serge N.

doi:10.1016/0141-8130(80)90045-8

Cited by 53 publications

(12 citation statements)

References 23 publications

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“…It is important to notice that for oxy-HbGp at pH 10.0 no contribution from the whole protein around 58 S [18] is observed. This suggests that alkaline pH 10.0 leads to complete oligomeric dissociation of HbGp into smaller M subunits as suggested previously also by gel filtration and SAXS experiments [19][20][21]28]. Another interesting observation from data shown in the Table 2 is that the percentage contribution of different species for HbGp at pH 10 is quite similar within experimental error, independently of the wavelength used for monitoring of absorbance: it is 25 ± 10% for the monomer, 30 ± 10% for dimers of subunit d and linkers, 36 ± 5% for trimers abc, and 9 ± 7% for tetramers abcd.…”

Section: Resultsmentioning

confidence: 52%

“…These heme-containing subunits form monomers of 16 kDa (d) and disulphide bound heterotrimers of 51-52 kDa (abc), linked by non-heme structures (24)(25)(26)(27)(28)(29)(30)(31)(32), named linkers (L) [3,5,7]. Recent partial characterization of M of HbGp by matrix assisted laser desorption time-of-flight mass spectrometry (MALDI-TOF-MS) confirmed the similarity of its subunits to those of orthologous proteins of this class, mentioned above [8].…”

Section: Introductionmentioning

confidence: 60%

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Molecular masses and sedimentation coefficients of extracellular hemoglobin of Glossoscolex paulistus: Alkaline oligomeric dissociation

Carvalho

Santiago

Borges

et al. 2011

International Journal of Biological Macromolecules

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Section: Resultsmentioning

confidence: 52%

Section: Introductionmentioning

confidence: 60%

Molecular masses and sedimentation coefficients of extracellular hemoglobin of Glossoscolex paulistus: Alkaline oligomeric dissociation

Carvalho

Santiago

Borges

et al. 2011

International Journal of Biological Macromolecules

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“…The same observation was made several years later by Boekema and van Heel (1989), when using electron microscopy with a resolution of 20 A . An analysis using small-angle X-ray scattering of Lumbricus terrestris EHb -the most extensively studied molecule -revealed the presence of a central subunit that was not visible with TEM (Pilz, Schwarz, and Vinogradov, 1980), in contrast to Tylorrynchus heterochaetus where the subunit was visible (Pilz et al, 1988). The same conclusion was drawn in the case of Arenicola marina and Glossoscolex paulistus EHbs (Wilhelm, Pilz, and Vinogradov, 1980;El Idrissi Slitine, Torriani, and Vachette, 1990).…”

Section: Architecturesupporting

confidence: 48%

Extracellular Hemoglobins from Annelids, and their Potential Use in Biotechnology

Zal

Rousselot

2014

Outstanding Marine Molecules

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“…3. This yielded a value of 12.7 Ϯ 2.8 nm (n ϭ 22) for R G which is 13% higher than the more precise value of 11.2 Ϯ 0.1 nm determined by low angle x-ray scattering (55). Somewhat higher accuracy may be possible with careful redetermination of the normalization factors for each detector, particularly at low angles (see "Materials and Methods") performed at the same time as experiments such as those shown in Fig.…”

Section: Hemoglobin Of L Terrestris Molecular Weightmentioning

confidence: 85%

Assembly of the Gigantic Hemoglobin of the Earthworm Lumbricus terrestris

Zhu

Ownby

Riggs

et al. 1996

Journal of Biological Chemistry

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The extracellular hemoglobin of the earthworm Lumbricus terrestris has four major kinds of O 2 -binding chains: a, b, and c (forming a disulfide-linked trimer), and chain d. Non-heme, non-globin structural chains, "linkers," are also present. Light-scattering techniques have been used to show that the ferrous CO-saturated abc trimer and chain d form an (abcd) 4 complex of 285 kDa at neutral pH. Formation of the full-sized 4-MDa molecule requires the addition of linker chains in the proportion of two linkers per (abcd) 4 and occurs much more rapidly in the presence of 10 mM calcium. This stoichiometry is supported not only by direct quantitative analysis of the intact hemoglobin but also by the fact that the addition of 50% of the proposed stoichiometric quantity of linkers results in the conversion of 50% of the (abcd) 4 to full-sized molecules. Isolated COsaturated abc trimers self-associate to (abc) 2 and higher aggregates up to an apparent limit of (abc) 10 ϳ550 kDa. The extracellular hemoglobins of annelids were first shown by Svedberg and Eriksson (1) to be gigantic molecules of at least 3 MDa in molecular mass. Although several subsequent studies gave molecular masses close to 4 MDa or higher for the Hb of Lumbricus terrestris and related species (2-4), recent studies by scanning transmission electron microscopy (5) have suggested masses of 3.5-3.6 MDa, and early light-scattering measurements suggested even lower values (6, 7). A possible reason for this great variation in reported molecular masses is oxidation which has been shown to cause extensive dissociation of many annelid Hbs (8 -10). Goss et al. (9) used light scattering to show that oxidation of L. terrestris Hb at neutral pH caused a large drop in molecular weight. Similarly, Ascoli et al. (10) found that oxidation of the similar Hb from Octolasium complanatum 1 caused hemichrome formation and dissociation outside of a narrow range near pH 7.The stoichiometry has also been uncertain. The Hb of L. terrestris has four major kinds of O 2 -binding globin chains: a, b, and c (forming the disulfide-linked abc trimer) and chain d, together with non-heme structural chains, "linkers," designated L. Kapp et al. (11) and Vinogradov et al. (12) have reported that linkers comprise 33-36% of the total mass, but this conclusion was based largely on the staining of SDS-gels with Coomassie Blue. This dye, however, binds to different proteins to quite different extents (13,14). Ownby et al. (15) redetermined the stoichiometry by reverse-phase high performance liquid chromatography (HPLC).2 The weight proportions of linkers were found to be approximately 16.4% by two independent procedures: the integrated absorption of the HPLC peaks and amino acid analysis of these peaks. These results led to the conclusion that the overall stoichiometry is (abcd) 2 L. This stoichiometry is further supported by the results of SDS-capillary gel electrophoresis monitored at 214 nm (see our companion study (16)).The goals of the present experiments are to redetermine the molecu...

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Small-angle X-ray studies of Lumbricus terrestris haemoglobin

Cited by 53 publications

References 23 publications

Molecular masses and sedimentation coefficients of extracellular hemoglobin of Glossoscolex paulistus: Alkaline oligomeric dissociation

Molecular masses and sedimentation coefficients of extracellular hemoglobin of Glossoscolex paulistus: Alkaline oligomeric dissociation

Extracellular Hemoglobins from Annelids, and their Potential Use in Biotechnology

Assembly of the Gigantic Hemoglobin of the Earthworm Lumbricus terrestris

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