title: Crystallization and preliminary structural analysis of the giant haemoglobin from Glossoscolex paulistus at 3.2 Å.
abstract: Glossoscolex paulistus is a free-living earthworm encountered in south-east Brazil. Its oxygen transport requirements are undertaken by a giant extracellular haemoglobin, or erythrocruorin (HbGp), which has an approximate molecular mass of 3.6 MDa and, by analogy with its homologue from Lumbricus terrestris (HbLt), is believed to be composed of a total of 180 polypeptide chains. In the present work the full 3.6 MDa particle in its cyanomet state was purified and crystallized using sodium citrate or PEG8000 as precipitant. The crystals contain one-quarter of the full particle in the asymmetric unit of the I222 cell and have parameters of a = 270.8 Å, b = 320.3 Å and c = 332.4 Å. Diffraction data were collected to 3.15 Å using synchrotron radiation on beamline X29A at the Brookhaven National Laboratory and represent the highest resolution data described to date for similar erythrocruorins. The structure was solved by molecular replacement using a search model corresponding to one-twelfth of its homologue from HbLt. This revealed that HbGp belongs to the type I class of erythrocruorins and provided an interpretable initial electron density map in which many features including the haem groups and disulfide bonds could be identified.
authors.affiliation: Centro de Biotecnologia Molecular Estrutural, Instituto de Fisica de São Carlos, Universidade de São Paulo, São Carlos-SP, CEP 13566-590, Brazil. bachega@ursa.if.sc.usp.br
authors.forename: J F R
authors.initials: JF
authors.lastname: Bachega
authors.forename: L
authors.initials: L
authors.lastname: Bleicher
authors.forename: E R
authors.initials: ER
authors.lastname: Horjales
authors.forename: P S
authors.initials: PS
authors.lastname: Santiago
authors.forename: R C
authors.initials: RC
authors.lastname: Garratt
authors.forename: M
authors.initials: M
authors.lastname: Tabak
mesh_terms: D000818:Animals; D000845:Annelida; D003460:Crystallization / Q000379:methods; D004902:Erythrocruorins / Q000737:chemistry*; D014961:X-Ray Diffraction
chemical_list: D004902:Erythrocruorins