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Three-Dimensional Immunoelectron Microscopy of Scorpion Hemocyanin Labeled with a Monoclonal Fab Fragment
Authors:Nicolas Boisset  Michael Radermacher  Robert Grassucci  Jean-Christophe Taveau  Weiping Liu  Josette Lamy  Joachim Frank  Jean N Lamy
Abstract:An immunocomplex of the 4 × 6-meric hemocyanin of the scorpion Androctonus australis with the monoclonal Fab fragment L104 was reconstructed from electron micrographs of a negatively stained specimen, using the double-carbon-layer technique. The resulting structure enables a clear visualization of the Fab fragments bound to the four copies of the Aa6 subunit and directly confirms a previous localization of the L104 epitope deduced from two-dimensional image processing. Despite a strong flattening effect produced by the negative-staining technique the orientations of the Fab fragments are well characterized. Moreover, the observation of a central hole within the elbow bends of the Fab fragments provides information about the disposition of the Fabs around their main axis.
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