Biochemical and electrophysiological characterization of N-glycans on NMDA receptor subunits

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216208%3A11310%2F16%3A10361682" target="_blank" >RIV/00216208:11310/16:10361682 - isvavai.cz</a>

Alternative codes found

RIV/67985823:_____/16:00463104

Result on the web

<a href="http://dx.doi.org/10.1111/jnc.13679" target="_blank" >http://dx.doi.org/10.1111/jnc.13679</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1111/jnc.13679" target="_blank" >10.1111/jnc.13679</a>

Result language

angličtina

Original language name

Biochemical and electrophysiological characterization of N-glycans on NMDA receptor subunits

Original language description

In mammals, excitatory synapses contain two major types of ionotropic glutamate receptors: -amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors and N-methyl-d-aspartate receptors (NMDARs). Both receptor types are comprised of several subunits that are post-translationally modified by N-glycosylation. However, the precise N-glycans that are attached to these receptor types are largely unknown. Here, we used biochemistry to confirm that native NMDARs are extensively N-glycosylated; moreover, we found that the NMDAR GluN2B subunit differs from GluN1 subunits with respect to endoglycosidase H sensitivity. Next, we used a complete panel of lectins to determine the glycan composition of NMDARs in both cerebellar tissue and cultured cerebellar granule cells. Our experiments identified 23 lectins that pulled down both the GluN1 and GluN2B NMDAR subunits. We then performed an electrophysiological analysis using representative lectins and found that pre-incubating cerebellar granule cells with the AAL, WGA, or ConA alters the receptor&apos;s biophysical properties; this lectin-mediated effect was eliminated when the cells were deglycosylated with peptide-N-glycosidase F. Similar lectin-mediated effects were observed using HEK293 cells that express recombinant GluN1/GluN2B receptors. Finally, using mutant recombinant GluN subunits expressed in HEK293 cells, we found that 11 out of 12 predicted N-glycosylation sites in GluN1 and 7 out of 7 N-glycosylation sites in GluN2B are occupied by N-glycans. These data provide new insight into the role that N-glycosylation plays in regulating the function of NMDA receptors in the central nervous system. All animal experiments were performed in accordance with relevant institutional ethics guidelines and regulations with respect to protecting animal welfare.

Czech name

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Czech description

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Type

J_imp - Article in a specialist periodical, which is included in the Web of Science database

CEP classification

—

OECD FORD branch

10601 - Cell biology

Project

—

Continuities

S - Specificky vyzkum na vysokych skolach

Publication year

2016

Confidentiality

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Name of the periodical

Journal of Neurochemistry

ISSN

0022-3042

e-ISSN

—

Volume of the periodical

138

Issue of the periodical within the volume

4

Country of publishing house

GB - UNITED KINGDOM

Number of pages

11

Pages from-to

546-556

UT code for WoS article

000382507300006

EID of the result in the Scopus database

2-s2.0-84981165843