Preparation of the polypeptides of the giant hemoglobin of Travisia japonica in carbon monoxy form

Investigation by Electrospray Ionization Mass Spectrometry of the Extracellular Hemoglobin from the Polychaete Annelid Alvinella pompejana: An Unusual Hexagonal Bilayer Hemoglobin

“…While V1 Hb possesses linker chains and shows a quaternary structure typical of HBL Hbs, the smaller Hbs V2 and C1 (M r ≈ 400 kDa) are devoid of linker chains and, although they contain roughly the same globin chains as V1, they do not form HBL structures (Suzuki et al, 1989;Zal et al, 1996a,b). An exception to this rule is the HBL Hb of the polychaete TraVisia japonica which reportedly consists of only five polypeptide chains containing heme with molecular weights in the range 14 000-18 000 Da (Fushitani et al, 1982). In AlVinella Hb, considering the potentially high temperatures that this species may encounter (Chevaldonne ´et al, 1992), the high proportion of linker chains may explain the thermostability of the molecule.…”

Extracellular Hemoglobins from Annelids, and their Potential Use in Biotechnology

“…When Suzuki et al (1983) were able to isolate (by HPLC) only six heme-components from the EHb of Perinereis brevicirris, their result was in agreement with that of Fushitani et al (1982a). Subsequently, all of these results were challenged, and following the determination of the primary sequences of three globin chains involved in the trimer subunit of Lumbricus EHb, Fushitani and Riggs (1988) recalculated the heme percentage.…”

“…However, the scientific community found this result very difficult to accept, and both Antonini and Chiancone (1977) and Garlick (1980) concluded that these results may have been due to purification artifacts. Subsequently, the absence of stoichiometry was admitted without real proof, although Fushitani et al (1982a) rejected this hypothesis using high-performance liquid chromatography (HPLC). In fact, these authors showed that the polychete EHb from Travisia japonica was assembled from five components with M r -values ranging between 14 and 18 kDa, each of which contained one heme, although the minimal molecular weight was about 23 000 Da (Fushitani et al, 1982a;Fushitani et al, 1982b;Fushitani, Morimoto, and Ochi, 1983) (Table 16.4).…”

Principles in the assembly of annelid erythrocruorins

“…Polypeptide chain separations under denaturing conditions generally show 2-4 bands in the 12-19,000 d range (23,26,28,37,(44)(45)(46) but higher molecular weight components are also present in most cases. Interchain disulfide linkages (23, 44, 45) and other aggregation properties tend to prevent complete separation.…”

“…Recently, progress has been made in the chromatographic separation of native functional heme-containing subunits in the presence of dithiothreitol (46)(47)(48). In these studies, nearly all of the protein is found in 18,000-d components that each contain heme.…”