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Three-dimensional reconstruction of native Androctonus australis hemocyanin
Authors:N Boisset  J C Taveau  J Lamy  T Wagenknecht  M Radermacher  J Frank
Institution:Laboratoire de Biochimie Fondamentale and URA, 1334 CNRS, Tours, France.
Abstract:A sample of native 4 x 6-meric hemocyanin of Androctonus australis was negatively stained with the double-layer technique, and was observed by transmission electron microscopy under low-dose conditions with a 50 degree and 0 degree tilt. The three-dimensional reconstruction method from "Single-exposure, random conical tilt series" was then applied. Independent three-dimensional reconstructions were obtained from the top, side and 45 degree views. Despite a pronounced flattening effect, presumably due to the specimen preparation technique, the positions of the 24 subunits composing the oligomer were unequivocally determined. This experiment definitely solves the problem of the architectural organization of the subunits in the cheliceratan 4 x 6-meric hemocyanins. Moreover, distinction between the flip and flop faces and an attenuated rocking effect were observed.
Keywords:
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