Dryad-Lohr-etal-westerns

These are all of the original images of the western blots in this manuscript. They are in a powerpoint file

Dryad-Lohr-etal-excelFSCV

This is an excel spreadsheet that shows all of the raw data collected for the fast scan cyclic voltammetry experiments. Tabs are used to delineate the various experiments

Conserved effects of the Pro552Arg mutation across GluN2 subunits.

<p><b>A</b>,<b>B</b>, Composite concentration-response curves of glutamate in the presence of 100 μM glycine (<b>A</b>) and glycine in the presence of 100 μM glutamate (<b>B</b>) for human GluN1-P557R/GluN2A, GluN1-P557R/GluN2B, GluN1/GluN2B-P553R, and rat GluN1/GluN2C-P550R, and GluN1/GluN2D-P577R. The graph legends refer to GluN1 as N1 and GluN2 as N2. Fitted EC₅₀ values are summarized in Tables <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1006536#pgen.1006536.t003" target="_blank">3</a> and <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1006536#pgen.1006536.t005" target="_blank">5</a>. <b>C</b>,<b>D</b>, human GluN1-P557R/GluN2A significantly prolongs deactivation time course after removal of glutamate (<b>C</b>) or removal of glycine (<b>D</b>) on transfected HEK293 cells, but does not slow the rise time when the receptors were activated by the agonists. <b>E</b>, GluN1/GluN2B-P553R significantly slows rise time and prolongs deactivation time course. Fitted parameters describing the response time course are given in <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1006536#pgen.1006536.t006" target="_blank">Table 6</a>.</p

Agonist potency of GluN2A-P552R equivalent mutations in GluN2B, GluN2C, and GluN2D.

<p>Agonist potency of GluN2A-P552R equivalent mutations in GluN2B, GluN2C, and GluN2D.</p

Analysis of genetic variation on S1-M1 linker in <i>GRIN1</i>, <i>GRIN2A</i>, <i>and GRIN2B</i>.

<p>Analysis of genetic variation on S1-M1 linker in <i>GRIN1</i>, <i>GRIN2A</i>, <i>and GRIN2B</i>.</p

Patients’ and mutations’ information.

<p>Patients’ and mutations’ information.</p

Pharmacology and response time course for substitution of GluN2A-ProP552 with Ala, Gly, Lys, Gln, Ile, or Leu.

<p>Pharmacology and response time course for substitution of GluN2A-ProP552 with Ala, Gly, Lys, Gln, Ile, or Leu.</p

Summary of pharmacological and functional properties of pre-M1 mutations.

<p>Summary of pharmacological and functional properties of pre-M1 mutations.</p

Potential interaction between the pre-M1 and M3 helices.

<p><b>A</b>,<b>B</b>, Ribbon structures of the GluN1/GluN2A (<b><i>A</i></b>) and GluN1/GluN2B (<b><i>B</i></b>) receptors without the amino terminal domain is shown. GluN1 is tan and GluN2 is light blue; regions with an OE-ratio below the 5^th percentile are colored purple, and indicate the regions under the strongest purifying selection. <b>C</b>, Side and top down view of the pore forming elements M1, M3, M4 in GluN1/GluN2A receptors colored as in (<b><i>A</i></b>), with regions of purifying selection shown in purple. <b>D</b>, Expanded view of the pre-M1 helix for GluN1 (<i>left panel</i>) and for GluN2A (<i>right panel</i>).</p

Assessment of alternative Pro552 substitutions in GluN2A.

<p><b>A,B</b>, the composite glutamate (in the presence of 100 μM glycine) and glycine (in the presence of 100 μM glutamate) concentration-response curves of GluN2A- P552A, P552G, P552I, P552K, P552Q, P552L constructs. Error bars are SEM and shown when larger than symbol. <b>C,D,E</b>, The response time courses are shown of GluN1/GluN2A(P552K), GluN1/GluN2A(P552G), and GluN1/GluN2A(P552L) receptors activated by rapid application of 100 μM glutamate; 100 μM glycine was present in all solutions. For panel <b>D</b> the rise time is expanded as an inset. The data (mean ± SEM) are given in <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1006536#pgen.1006536.t007" target="_blank">Table 7</a>.</p