title: Helix-helix interactions between homo- and heterodimeric gamma-carboxyglutamate-containing conantokin peptides and their derivatives.
abstract: The conantokins are a family of small, naturally occurring gamma-carboxyglutamate (Gla)-rich peptides that specifically antagonize the N-methyl-D-aspartate (NMDA) subtype of ionotropic glutamate receptor. One member of this family, conantokin-G (con-G), undergoes Ca(2+)-mediated self-assembly to form an antiparallel helical dimer. Subunit interactions in this complex are incumbent upon intermolecular Ca(2+) bridging of Gla residues spaced at i, i + 4, i + 7, i + 11 intervals within the monomer. Herein, we further probe the molecular determinants governing such helix-helix interactions. Select variants were synthesized to evaluate the contributions of non-Gla residues to conantokin self-association. Con-G dimerization was shown to be exothermic and accompanied by positive heat capacity changes. Using positional Gla variants of conantokin-R (con-R), a non-dimerizing conantokin, i, i + 4, i + 7, i + 11 Gla spacing alone was shown to be insufficient for self-assembly. The Ca(2+)-dependent antiparallel heterodimerization of con-G and con-T(K7 gamma), two peptides that harbor optimal Gla spacing, was established. Last, the effects of covalently constrained con-G dipeptides on NMDA-evoked current in HEK293 cells expressing combinations of NR1a, NR1b, NR2A, and NR2B subunits of the NMDA receptor were investigated. The antiparallel dipeptide was unique in its ability to potentiate current at NR1a/2A receptors and, like monomeric con-G, was inhibitory at NR1a/2B and NR1b/2B combinations. In contrast, the parallel species was completely inactive at all subunit combinations tested. These results suggest that, under physiological Ca(2+) concentrations, equilibrium levels of con-G dimer most likely exist in an antiparallel orientation and exert effects on NMDA receptor activity that differ from the monomer.
authors.affiliation: Department of Chemistry and Biochemistry and the W.M. Keck Center for Transgene Research, University of Notre Dame, Notre Dame, Indiana 46556, USA.
authors.forename: Qiuyun
authors.initials: Q
authors.lastname: Dai
authors.forename: Zhenyu
authors.initials: Z
authors.lastname: Sheng
authors.forename: James H
authors.initials: JH
authors.lastname: Geiger
authors.forename: Francis J
authors.initials: FJ
authors.lastname: Castellino
authors.forename: Mary
authors.initials: M
authors.lastname: Prorok
mesh_terms: D015055:1-Carboxyglutamic Acid / Q000737:chemistry*; D002118:Calcium / Q000737:chemistry*; D002460:Cell Line; D020916:Conotoxins / Q000138:chemical synthesis / Q000737:chemistry* / Q000494:pharmacology; D019281:Dimerization; D006801:Humans; D008564:Membrane Potentials / Q000187:drug effects; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry* / Q000494:pharmacology; D017433:Protein Structure, Secondary
chemical_list: D020916:Conotoxins; D010455:Peptides; D015055:1-Carboxyglutamic Acid; C055407:conotoxin GV; D002118:Calcium