Quaternary structure, protein dynamics, and synaptic function of SAP97 controlled by L27 domain interactions.
Single-particle electron microscopy (EM) combined with biochemical measurements revealed the molecular shape of SAP97 and a monomer-dimer transition that depended on the N-terminal L27 domain. Overexpression of SAP97 drove GluR1 to synapses, potentiated AMPA receptor (AMPAR) excitatory postsynaptic currents (EPSCs), and occluded LTP. Synaptic potentiation and GluR1 delivery were dissociable by L27 domain mutants that inhibit multimerization of SAP97. Loss of potentiation was correlated with faster turnover of monomeric SAP97 mutants in dendritic spines. We propose that L27-mediated interactions of SAP97 with itself or other proteins regulate the synaptic delivery of AMPARs. RNAi knockdown of endogenous PSD-95 depleted surface GluR1 and impaired AMPA EPSCs. In contrast, RNAi knockdown of endogenous SAP97 reduced surface expression of both GluR1 and GluR2 and inhibited both AMPA and NMDA EPSCs. Thus SAP97 has a broader role than its close relative, PSD-95, in the maintenance of synaptic function.
Nakagawa
Terunaga
T
The Picower Center for Learning and Memory, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, MA 02139, USA.
Futai
Kensuke
K
Lashuel
Hilal A
HA
Lo
Irene
I
Okamoto
Kenichi
K
Walz
Thomas
T
Hayashi
Yasunori
Y
Sheng
Morgan
M
D048868:Adaptor Proteins, Signal Transducing; D000704:Analysis of Variance; D000818:Animals; D016367:Baculoviridae / Q000502:physiology; D001703:Biophysics / Q000379:methods; D001921:Brain / Q000166:cytology; D002478:Cells, Cultured; D002850:Chromatography, Gel / Q000379:methods; D049229:Dendritic Spines / Q000378:metabolism; D000074288:Discs Large Homolog 1 Protein; D000076122:Disks Large Homolog 4 Protein; D018690:Excitatory Amino Acid Agonists / Q000494:pharmacology; D018691:Excitatory Amino Acid Antagonists / Q000494:pharmacology; D019706:Excitatory Postsynaptic Potentials / Q000235:genetics; D005786:Gene Expression Regulation / Q000502:physiology; D005822:Genetic Vectors; D051528:Guanylate Kinases; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D066298:In Vitro Techniques; D007313:Insecta; D047908:Intracellular Signaling Peptides and Proteins; D016086:Lentivirus / Q000235:genetics; D017774:Long-Term Potentiation / Q000235:genetics; D008565:Membrane Proteins; D051379:Mice; D018613:Microscopy, Confocal / Q000379:methods; D008855:Microscopy, Electron, Scanning / Q000379:methods; D046032:Microscopy, Energy-Filtering Transmission Electron / Q000379:methods; D016296:Mutagenesis; D016202:N-Methylaspartate / Q000494:pharmacology; D009419:Nerve Tissue Proteins / Q000737:chemistry* / Q000378:metabolism / Q000502:physiology* / Q000648:ultrastructure; D009474:Neurons / Q000166:cytology / Q000502:physiology; D018408:Patch-Clamp Techniques / Q000379:methods; D017434:Protein Structure, Tertiary / Q000502:physiology*; D016372:RNA, Antisense / Q000378:metabolism; D034741:RNA, Small Interfering; D051381:Rats; D018091:Receptors, AMPA / Q000378:metabolism / Q000502:physiology; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment / Q000379:methods; D013569:Synapses / Q000502:physiology*; D009435:Synaptic Transmission / Q000502:physiology*; D013997:Time Factors; D014162:Transfection / Q000379:methods; D014633:Valine / Q000031:analogs & derivatives* / Q000494:pharmacology
D048868:Adaptor Proteins, Signal Transducing; D000074288:Discs Large Homolog 1 Protein; D000076122:Disks Large Homolog 4 Protein; D018690:Excitatory Amino Acid Agonists; D018691:Excitatory Amino Acid Antagonists; D051528:Guanylate Kinases; D047908:Intracellular Signaling Peptides and Proteins; D008565:Membrane Proteins; D016202:N-Methylaspartate; D009419:Nerve Tissue Proteins; D016372:RNA, Antisense; D034741:RNA, Small Interfering; D018091:Receptors, AMPA; D011993:Recombinant Fusion Proteins; D014633:Valine; C513314:DLG1 protein, human; C000614189:Dlg1 protein, mouse; C501119:Dlg1 protein, rat; C501447:Dlg4 protein, mouse; C501449:Dlg4 protein, rat; C033652:postsynaptic density proteins; C056639:2-amino-5-phosphopentanoic acid