an alternating glun1-2-1-2 subunit arrangement in mature nmda receptors成熟的门冬氨酸受体的交替glun1-2-1-2亚基的安排.pdfVIP

An Alternating GluN1-2-1-2 Subunit Arrangement in Mature NMDA Receptors 1,2,3. 1,2,3. 4 1,2,3 Morgane Riou , David Stroebel , J. Michael Edwardson , Pierre Paoletti * ´ 1 Ecole Normale Superieure, Institut de Biologie de l’ENS, IBENS, Paris, France, 2 Inserm, U1024, Paris, France, 3 CNRS, UMR 8197, Paris, France, 4 Department of Pharmacology, University of Cambridge, Cambridge, United Kingdom Abstract NMDA receptors (NMDARs) form glutamate-gated ion channels that play a critical role in CNS physiology and pathology. Together with AMPA and kainate receptors, NMDARs are known to operate as tetrameric complexes with four membrane- embedded subunits associating to form a single central ion-conducting pore. While AMPA and some kainate receptors can function as homomers, NMDARs are obligatory heteromers composed of homologous but distinct subunits, most usually of the GluN1 and GluN2 types. A fundamental structural feature of NMDARs, that of the subunit arrangement around the ion pore, is still controversial. Thus, in a typical NMDAR associating two GluN1 and two GluN2 subunits, there is evidence for both alternating 1/2/1/2 and non-alternating 1/1/2/2 arrangements. Here, using a combination of electrophysiological and cross-linking experiments, we provide evidence that functional GluN1/GluN2A receptors adopt the 1/2/1/2 arrangement in which like subunits are diagonal to one another. Moreover, based on the recent crystal structure of an AMPA receptor, we show that in the agonist-binding and pore regions, the GluN1 subunits occupy a ‘‘proximal’’ position, closer to the central axis of the channel pore than that of GluN2 subunits. Finally, results obtained w