Abstract
The postsynaptic density protein PSD-95 and related membrane-associated guanylate kinases are scaffolding proteins, whose modular interaction motifs organize protein complexes at cell junctions. The signature guanylate kinase domain (GK) contains elements of the protein's GMP-binding site but does not bind nucleotide. Instead, the GK domain has evolved from an enzyme to a protein-protein interaction motif. Here, we show that this canonical GMP-binding region interacts with microtubule-associated protein-1a (MAP1a) and we present a structural model. We determine the consensus GK-binding sequence in MAP1a and demonstrate that PSD-95 can use a similar interaction mode to bind diverse protein partners. Furthermore, we show that PSD-95 GK has adopted the conformational flexibility of the ancestral enzyme to bind its varied ligands, which suggests a mechanism of regulation.
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Acknowledgements
We thank J. Gross for space and advice, and M. Bowen, M. Daugherty, S. Carter, J. Flinders, Q. Justman, E. LaDow, S. Newmyer, S. Pintchovski, M. Pufall and R. Yu for critical comments on the manuscript. This work was supported by grants from the Deutsche Forschungsgemeinschaft (SFB 628), the European Union (HPRI-CT-2001-50028) and the Center for Biomolecular Magnetic Resonance at the University of Frankfurt. M.L.R. was a US National Science Foundation Graduate Research Fellow and was a recipient of a Boehringer-Ingelheim Fonds Travel Grant.
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Supplementary information
Supplementary Fig. 1
Titration of [15N]lysine-labeled SH3-GK with MAP1a peptide. (PDF 256 kb)
Supplementary Fig. 2
Peak assignment through mutation. (PDF 298 kb)
Supplementary Fig. 3
Orientation of MAP1a by paramagnetic relaxation enhancement. (PDF 131 kb)
Supplementary Fig. 4
[15N]tyrosine SH3-GK titration with MAP1a. (PDF 156 kb)
Supplementary Fig. 5
Top-scored models from docking simulations (PDF 1312 kb)
Supplementary Fig. 6
Analysis of matched cysteine mutations by fluorescence anisotropy. (PDF 135 kb)
Supplementary Fig. 7
Structural alignment of GK domains. (PDF 1029 kb)
Supplementary Fig. 8
Alignment of the SH3 and Hook domains of the neuronal MAGUK family. (PDF 350 kb)
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Reese, M., Dakoji, S., Bredt, D. et al. The guanylate kinase domain of the MAGUK PSD-95 binds dynamically to a conserved motif in MAP1a. Nat Struct Mol Biol 14, 155–163 (2007). https://doi.org/10.1038/nsmb1195
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DOI: https://doi.org/10.1038/nsmb1195
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