title: Immunoelectron microscopic localization of glutamyl-/ prolyl-tRNA synthetase within the eukaryotic multisynthetase complex.
abstract: A high molecular mass complex of aminoacyl-tRNA synthetases is readily isolated from a variety of eukaryotes. Although its composition is well characterized, knowledge of its structure and organization is still quite limited. This study uses antibodies directed against prolyl-tRNA synthetase for immunoelectron microscopic localization of the bifunctional glutamyl-/prolyl-tRNA synthetase. This is the first visualization of a specific site within the multisynthetase complex. Images of immunocomplexes are presented in the characteristic views of negatively stained multisynthetase complex from rabbit reticulocytes. As described in terms of a three domain working model of the structure, in "front" views of the particle and "intermediate" views, the primary antibody binding site is near the intersection between the "base" and one "arm." In "side" views, where the particle is rotated about its long axis, the binding site is near the midpoint. "Top" and "bottom" views, which appear as square projections, are also consistent with the central location of the binding site. These data place the glutamyl-/prolyl-tRNA synthetase polypeptide in a defined area of the particle, which encompasses portions of two domains, yet is consistent with the previous structural model.
authors.affiliation: Department of Biochemistry, The University of Mississippi Medical Center, Jackson, Mississippi 39216-4505, USA. mnorcum@biochem.umsmed.edu
authors.forename: M T
authors.initials: MT
authors.lastname: Norcum
authors.forename: J D
authors.initials: JD
authors.lastname: Dignam
mesh_terms: D000604:Amino Acyl-tRNA Synthetases / Q000378:metabolism*; D000818:Animals; D000918:Antibody Specificity; D005975:Glutamate-tRNA Ligase / Q000276:immunology / Q000378:metabolism*; D016253:Microscopy, Immunoelectron; D009097:Multienzyme Complexes / Q000378:metabolism*; D011817:Rabbits
chemical_list: D009097:Multienzyme Complexes; D000604:Amino Acyl-tRNA Synthetases; C100134:prolyl T RNA synthetase; D005975:Glutamate-tRNA Ligase