Mass spectrometry identifies LGI1-interacting proteins that are involved in synaptic vesicle function in the human brain.
The LGI1 gene has been shown to predispose to epilepsy and influence cell invasion in glioma cells. To identify proteins that interact with LGI1 and gain a better understanding of its function, we have used co-immunoprecipitation (co-IP) of a secreted green fluorescent protein-tagged LGI1 protein combined with mass spectrometry to identify interacting partners from lysates prepared from human subcortical white matter. Proteins were recovered from polyacrylamide gels and analyzed using liquid chromatography coupled to tandem mass spectrometry. This analysis identified a range of proteins, but in particular synaptotagmin, synaptophysin, and syntaxin 1A. Each of these proteins is found associated with synaptic vesicles. These interactions were confirmed independently by co-IP and Western blotting and implicate LGI1 in synapse biology in neurons. Other vesicle-related proteins that were recovered by co-IP include clathrin heavy chain 1, syntaxin binding protein 1, and a disintegrin and metalloprotease 23. These observations support a role for LGI1 in synapse vesicle function in neurons.
Kunapuli
Padmaja
P
Department of Cell Stress Biology, Roswell Park Cancer Institute, Buffalo, NY 14263, USA.
Jang
Geeng-Fu
GF
Kazim
Latif
L
Cowell
John K
JK
D000595:Amino Acid Sequence; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D013058:Mass Spectrometry / Q000379:methods*; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism*; D011506:Proteins / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D013572:Synaptic Vesicles / Q000378:metabolism*
D047908:Intracellular Signaling Peptides and Proteins; C116499:LGI1 protein, human; D009419:Nerve Tissue Proteins; D011506:Proteins; D011993:Recombinant Fusion Proteins