콘텐츠로 건너뛰기
Merck
  • Anti-NMDA receptor encephalitis antibody binding is dependent on amino acid identity of a small region within the GluN1 amino terminal domain.

Anti-NMDA receptor encephalitis antibody binding is dependent on amino acid identity of a small region within the GluN1 amino terminal domain.

The Journal of neuroscience : the official journal of the Society for Neuroscience (2012-08-10)
Amy J Gleichman, Lynn A Spruce, Josep Dalmau, Steven H Seeholzer, David R Lynch
초록

Anti-NMDA receptor (NMDAR) encephalitis is a newly identified autoimmune disorder that targets NMDARs, causing severe neurological symptoms including hallucinations, psychosis, and seizures, and may result in death (Dalmau et al., 2008). However, the exact epitope to which these antibodies bind is unknown. A clearly defined antigenic region could provide more precise testing, allow for comparison of immunogenicity between patients to explore potential clinically relevant variations, elucidate the functional effects of antibodies, and make patients' antibodies a more effective tool with which to study NMDAR function. Here, we use human CSF to explore the antigenic region of the NMDAR. We created a series of mutants within the amino terminal domain of GluN1 that change patient antibody binding in transfected cells in stereotyped ways. These mutants demonstrate that the N368/G369 region of GluN1 is crucial for the creation of immunoreactivity. Mass spectrometry experiments show that N368 is glycosylated in transfected cells and rat brain regions; however, this glycosylation is not directly required for epitope formation. Mutations of residues N368/G369 change the closed time of the receptor in single channel recordings; more frequent channel openings correlates with the degree of antibody staining, and acute antibody exposure prolongs open time of the receptor. The staining pattern of mutant receptors is similar across subgroups of patients, indicating consistent immunogenicity, although we have identified one region that has a variable role in epitope formation. These findings provide tools for detailed comparison of antibodies across patients and suggest an interaction between antibody binding and channel function.

MATERIALS
제품 번호
브랜드
제품 설명

Sigma-Aldrich
Anti-GFP, N-terminal antibody produced in rabbit, ~1.0 mg/mL, affinity isolated antibody, buffered aqueous solution
Sigma-Aldrich
Anti-NR1 Antibody, CT, Upstate®, from mouse
Sigma-Aldrich
Anti-NMDAR1 Antibody, rabbit monoclonal, culture supernatant, clone 1.17.2.6, Chemicon®
Sigma-Aldrich
Anti-Actin, N-terminal antibody produced in rabbit, ~0.5 mg/mL, affinity isolated antibody, buffered aqueous solution