Revisiting the structure of Alvinella pompejana hemoglobin at 20A resolution by cryoelectron microscopy.
The hemoglobin of the polychaete worm Alvinella pompejana was reconstructed at 20A resolution from frozen-hydrated samples observed by electron microscopy according to the random conical tilt series method. This three-dimensional reconstruction was mirror-inverted with respect to a previous volume published by de Haas et al. in 1996. In order to explain this handedness discrepancy, various 3D reconstructions using different reference volumes were carried out showing that the choice of the first volume was the keystone during the refinement process. The 3D reconstruction volume of A. pompejana Hb presented structural features characteristic of annelid Hbs with two hexagonal layers each comprising six hollow globular subassemblies and a complex of non-heme linker chains. Moreover, the eclipsed conformation of the two hexagonal layers and a HGS architecture similar to that described for Arenicola marina Hb led to the conclusion that A. pompejana Hb belonged to the architectural type II according to the definition of Jouan et al. (2001). A comparison between this cryo-EM volume and X-ray crystallography density maps of Lumbricus terrestris type-I Hb (Royer et al., 2000) showed that the triple stranded coiled coil structures of linker chains were different. Based on this observation, a model was proposed to explain the eclipsed conformation of the two hexagonal layers of type-II Hbs.
Jouan
Ludovic
L
Laboratoire des Protéines Complexes, J. E. 2320, Université de Tours. 2, bis Boulevard Tonnellé, F-37032 Tours Cedex, France.
Marco
Sergio
S
Taveau
Jean-Christophe
JC
D000818:Animals; D000845:Annelida / Q000378:metabolism*; D020285:Cryoelectron Microscopy / Q000379:methods*; D018360:Crystallography, X-Ray; D006418:Heme / Q000737:chemistry; D006454:Hemoglobins / Q000737:chemistry* / Q000648:ultrastructure*; D007091:Image Processing, Computer-Assisted; D021621:Imaging, Three-Dimensional; D008958:Models, Molecular; D012984:Software
D006454:Hemoglobins; D006418:Heme