Mass Spectrometic Composition, Molecular Mass and Oxygen Binding ofMacrobdella decoraHemoglobin and its Tetramer and Monomer Subunits

Weber, Roy E.; Malte, Hans; Braswell, Emory H.; Oliver, R.; Green, B.N.; Sharma, Pawan K.; Kuchumov, Askar R.; Vinogradov, Serge N.

doi:10.1006/jmbi.1995.0466

Cited by 43 publications

(70 citation statements)

References 55 publications

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“…The subunit composition of Nephelopsis Hb is in agreement with those of the leeches Macrobdella decora [15] and Haemopis sanguisuga [16] and is also found for the deep-sea hydrothermal vent polychaete, Riftia pachyptila [17]. These results illustrate the potency of the MaxEnt deconvolution software in disentangling the multiply charged data produced from complex protein mixtures by ESI-MS.…”

Section: Resultssupporting

confidence: 76%

An electrospray ionization mass spectrometric study of the subunit structure of the giant hemoglobin from the leech Nephelopsis oscura

Green

Vinogradov

2004

J. Am. Soc. Mass Spectrom.

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The subunit structure of the giant, extracellular hexagonal bilayer (HBL) hemoglobin (Hb) from the leech Nephelopsis oscura was investigated by electrospray ionization mass spectrometry (ESI-MS) employing a maximum entropy deconvolution of its complex, multiply charged ESI spectra. The denatured unreduced Hb consisted of three monomer globin chains These giant complexes were found to have masses of ϳ3500 kDa and to consist of two types of polypeptide chains: heme-containing 16 -17 kDa globin chains and non-globin, linker chains of 24 -32 kDa containing 9 to 12 cysteines [4]. The amino acid sequences of the globin chains are clearly related to vertebrate and invertebrate globins [5] and they form disulfide-bonded dimer, trimer and tetramer subunits as well as noncovalent subassemblies, ranging from dimer to dodecamer [4].Over the last decade, ESI-MS has been shown to be the method of choice for the characterization of proteins [6,7] by providing highly accurate masses. In particular, in the case of heteromultimeric protein complexes, as exemplified by the HBL Hbs, ESI-MS provides an exquisitely accurate enumeration of the constituent chains and covalently bonded subunits, as well as a quantitative determination of free and disulfidebonded Cys residues [8]. Furthermore, the combination of nanoflow electrospray and time-of-flight (TOF) mass analyzers has recently allowed the investigation of the quaternary structure of large noncovalent protein complexes [9].We describe here the results of ESI-MS and ESI-TOF-MS studies of the polypeptide chain and subunit composition and noncovalent subassemblies of the HBL Hb from the leech Nephelopsis oscura.

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Section: Resultssupporting

confidence: 76%

An electrospray ionization mass spectrometric study of the subunit structure of the giant hemoglobin from the leech Nephelopsis oscura

Green

Vinogradov

2004

J. Am. Soc. Mass Spectrom.

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“…3B) and we found that the glycosylated isoforms a1-a5 differ in mass by one or more hexose residue(s) (162.1 Da). The difference in mass between chain a5 and chain a (2680.3 Da) is, within experimental error, the mass of the glycan residue (HexNAc) 2 (Hex) 14 (2676.40 Da). Note that ESI-MS cannot distinguish between monosaccharide residues because they have identical masses.…”

Section: Polypeptide Chains Composition Determined By Esi-ms-mentioning

confidence: 80%

“…More recently, a more accurate technique, ESI-MS, has been successfully applied to some HBL Hbs allowing the determination of the complete polypeptide chain compositions of these Hbs (13)(14)(15). They consist of 4 to 6 globin chains in the range 16,000 -18,000 Da and 3-4 linker chains in the range 24,000 -27,000 Da.…”

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confidence: 99%

The Multi-hemoglobin System of the Hydrothermal Vent Tube Worm Riftia pachyptila

Zal

Lallier

Green³

et al. 1996

Journal of Biological Chemistry

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The deep-sea tube worm Riftia pachyptila Jones possesses a complex of three extracellular Hbs: two in the vascular compartment, V1 (ϳ3500 kDa) and V2 (ϳ400 kDa), and one in the coelomic cavity, C1 (ϳ400 kDa). These native Hbs, their dissociation products and derivatives were subjected to electrospray ionization mass spectrometry (ESI-MS). The data were analyzed by the maximum entropy deconvolution system. We identified three groups of peaks for V1 Hb, at ϳ16, 23-27, and 30 kDa, corresponding to ( Several models have been proposed for the quaternary structure of annelid hexagonal bilayer hemoglobins (Hbs) 1 (1-5) and vestimentifera Hbs (6) but no definitive agreement exists. However, it is well established that HBL Hbs have a hexagonal symmetry and consist of two types of chains, globin-chains (ϳ17,000 Da) accounting for approximately 70% of total mass and heme-deficient linker chains (ϳ24,000 -32,000 Da) necessary for assemblage into the HBL structure (7). A detailed understanding of this molecular structure requires the knowledge of: (i) the molecular mass of the native molecule; (ii) the number and proportions of all constitutive polypeptide chains; (iii) the number and relation between subunits; and (iv) the determination of linker proportions. In a companion study (8) we have confirmed that Riftia pachyptila (Jones), a vestimentiferan living around deep-sea hydrothermal vents (9 -11), possesses three hemoglobins, two of them dissolved in the vascular blood (V1 and V2), and one in the coelomic fluid (C1). Their molecular weights have been determined by scanning transmission electron microscopy mass mapping (STEM) and by multi-angle laser light scattering (MALLS). Both methods yielded approximately the same molecular weights with mass significantly higher than the literature data for V1. V1, V2, and C1 had M r of 3396 Ϯ 540 ϫ 10 3

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“…This equation was fit to the data in the form log (Y/1 Ϫ Y) versus log P (end-weighting), and parameters were estimated using the curve-fitting procedure described by Weber et al (47). In separate analyses, values of q were estimated from the data or were fixed at 4, as applies to tetrameric Hb.…”

Section: Methodsmentioning

confidence: 99%

Gene Duplication and the Evolution of Hemoglobin Isoform Differentiation in Birds

Grispo

Natarajan

Projecto-Garcia

et al. 2012

Journal of Biological Chemistry

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Background:The functional significance of hemoglobin heterogeneity remains a mystery. Results: In adult birds, the HbD isoform (related to embryonic hemoglobin) exhibits distinct oxygenation properties relative to the major HbA isoform. Conclusion: Substitutions that distinguish HbD from HbA are not shared with embryonic hemoglobin. Significance: Differences between isoforms stem from derived (nonancestral) changes in duplicated genes, not from the retention of an ancestral condition.

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Mass Spectrometic Composition, Molecular Mass and Oxygen Binding ofMacrobdella decoraHemoglobin and its Tetramer and Monomer Subunits

Cited by 43 publications

References 55 publications

An electrospray ionization mass spectrometric study of the subunit structure of the giant hemoglobin from the leech Nephelopsis oscura

An electrospray ionization mass spectrometric study of the subunit structure of the giant hemoglobin from the leech Nephelopsis oscura

The Multi-hemoglobin System of the Hydrothermal Vent Tube Worm Riftia pachyptila

Gene Duplication and the Evolution of Hemoglobin Isoform Differentiation in Birds

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