Investigation by electrospray ionization mass spectrometry of the extracellular hemoglobin from the polychaete annelid Alvinella pompejana: an unusual hexagonal bilayer hemoglobin.
Alvinella pompejana inhabits deep-sea hydrothermal vent sites along the East-Pacific Rise, where it colonizes the walls of actively venting high-temperature chimneys. This worm is the most thermophilic metazoan known to date. In Alvinella, as in other alvinellids, oxygen transport is mainly achieved by an extracellular Hb dissolved in the vascular blood. This Hb has a molecular mass of 3833 +/- 14 kDa as revealed by multiangle laser light scattering (MALLS). Native and derivative Hb (reduced, carbamidomethylated, and deglycosylated) were analyzed by electrospray ionization mass spectrometry (ESI-MS). The data were processed by the maximum entropy deconvolution system (MaxEnt). We identified three groups of peaks for Alvinella Hb, at ca. 16, 23-26, and 50 kDa corresponding to (i) four monomeric globin chains, a1 (16 633.4), a2(16 532.4), a3 (16 419.6), and a4(16 348.9); (ii) four linker chains, L1-L4 (22 887. 1, 24 230.5, 26 233.6, and 25 974.4); and (iii) one disulfide-bonded trimer T (51 431.9) composed of globin chains b (16 477.5), c (16 916.1), and d (18 048.8). These Hbs were also subjected to SDS-PAGE analysis for comparative purposes. In addition, using the ESI-MS data we propose two alternative models for the quaternary structure of Alvinella's Hb.
Zal
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Equipe Ecophysiologie, UPMC-CNRS-INSU, Station Biologique, BP 74, 29682 Roscoff Cedex, France, and Micromass UK Ltd., Tudor Road, Altrincham, Cheshire WA14 5RZ, U.K. zal@lifesci.lscf.ucsf.edu
Green
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Lallier
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Toulmond
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D000818:Animals; D002241:Carbohydrates / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid; D003545:Cysteine / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D006454:Hemoglobins / Q000737:chemistry*; D007834:Lasers; D013058:Mass Spectrometry; D011077:Polychaeta; D011487:Protein Conformation; D012542:Scattering, Radiation
D002241:Carbohydrates; D006454:Hemoglobins; D003545:Cysteine