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. Author manuscript; available in PMC: 2011 Jun 8.
Published in final edited form as: J Neurosci. 2010 Dec 8;30(49):16437–16452. doi: 10.1523/JNEUROSCI.4478-10.2010

Figure 7. Calcium-induced modification of PICK1 structure requires the N-terminal acidic motif and is essential for LTD.

Figure 7

A, CD spectra of recombinant PICK1 in the absence (open circles) or presence (solid circles) of 1 mM calcium. B, Thermal denaturation of recombinant PICK1 monitored through the CD signal at 220 nm in the absence (open circles) or presence (solid circles) of 1 mM calcium. C, CD spectra of recombinant PICK1(D3N) in the absence (open triangles) or presence (solid triangles) of 1 mM calcium. D, Thermal denaturation of recombinant PICK1(D3N) in the absence (open triangles) or presence (solid triangles) of 1 mM calcium. E, Superposition of the CD spectra of recombinant WT PICK1 in the presence of 1 mM calcium (solid circles) with that of PICK1(D3N) in the absence of calcium (open triangles). F, Thermal denaturation of recombinant PICK1(Δ9) in the absence (open diamonds) or presence (solid diamonds) of 1 mM calcium. G–H, Schematics depicting the N-terminal domain mutants PICK1(Δ9) (G) and PICK1(D3N) (H). I–J, AMPAR EPSCs are not affected by replacement of PICK1 with GFP-PICK1(Δ9) (I; shPICK1 + GFP-PICK1(Δ9), −72.1±8.3 pA, uninfected: −74.3±12.4 pA, n=16 pairs) or GFP-PICK1(D3N) (J; shPICK1 + GFP-PICK1(D3N), −64.6±8.3 pA, uninfected: −65.7±8.4 pA, n=13 pairs). K–L, LTD is inhibited by replacement of endogenous PICK1 with GFP-PICK1(Δ9) (K) or GFP-PICK1(D3N) (L). K1, L1, Sample experiments demonstrating lack of LTD in a cell expressing shPICK1 + GFP-PICK1(Δ9) (K1) or a cell expressing shPICK1 + GFP-PICK1(D3N) (L1). K2, L2, Summary graphs demonstrating block of LTD in shPICK1 + GFP-PICK1(Δ9) infected neurons (K2; shPICK1 + GFP-PICK1(Δ9), 93±5%, n=16; uninfected: 46±7%, n=14) or shPICK1 + GFP-PICK1(D3N) infected neurons (L2; shPICK1 + GFP-PICK1(D3N), 102±11%, n=10; uninfected: 46±7%, n=14; the same uninfected control graph is shown in both panels because the PICK1(Δ9) and PICK1(D3N) mutants were analyzed in parallel and are therefore compared to a common set of interleaved controls).