Structural highlights
Function
[DLG4_RAT] Interacts with the cytoplasmic tail of NMDA receptor subunits and shaker-type potassium channels. Required for synaptic plasticity associated with NMDA receptor signaling. Overexpression or depletion of DLG4 changes the ratio of excitatory to inhibitory synapses in hippocampal neurons. May reduce the amplitude of ASIC3 acid-evoked currents by retaining the channel intracellularly. May regulate the intracellular trafficking of ADR1B.[1] [2]
Publication Abstract from PubMed
The impact of the incorporation of a non-natural amino acid (NNAA) onto protein structure, dynamics, and ligand binding has not been studied rigorously so far. NNAAs are regularly used to modify proteins post-translationally in vivo and in vitro using click-chemistry. Here, we present a structural characterization of the impact of the incorporation of azidohomoalanine (AZH) into the model protein domain PDZ3 using NMR spectroscopy and X-ray crystallography. The structure and dynamics of the apo state of the AZH-modified PDZ3 remains mostly unperturbed. Furthermore, the binding of two PDZ3 binding peptides are found to be unchanged by incorporation of AZH. We mapped the interface of the AZH-modified PDZ3 and an azulene-linked peptide for vibrational energy transfer studies using chemical shift perturbations and NOEs between the unlabelled azulene-linked peptide and the isotopically labelled protein. Co-crystallization and soaking failed for the peptide-bound holo-complex. NMR spectroscopy, however, al-lowed determination of the protein-ligand interface. While the incorpo-ration of AZH was minimally invasive for PDZ3, structural analysis of NNAA-modified proteins by the methodology presented here should be carried out to ensure structural integrity of the studied target.
The Impact of Azidohomoalanine Incorporation on Protein Structure and Ligand Binding.,Lehner F, Kudlinzki D, Richter C, Muller-Werkmeister H, Eberl K, Bredenbeck J, Schwalbe H, Silvers R Chembiochem. 2017 Sep 26. doi: 10.1002/cbic.201700437. PMID:28950050[3]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
References
- ↑ Hruska-Hageman AM, Benson CJ, Leonard AS, Price MP, Welsh MJ. PSD-95 and Lin-7b interact with acid-sensing ion channel-3 and have opposite effects on H+- gated current. J Biol Chem. 2004 Nov 5;279(45):46962-8. Epub 2004 Aug 17. PMID:15317815 doi:10.1074/jbc.M405874200
- ↑ Prange O, Wong TP, Gerrow K, Wang YT, El-Husseini A. A balance between excitatory and inhibitory synapses is controlled by PSD-95 and neuroligin. Proc Natl Acad Sci U S A. 2004 Sep 21;101(38):13915-20. Epub 2004 Sep 9. PMID:15358863 doi:10.1073/pnas.0405939101
- ↑ Lehner F, Kudlinzki D, Richter C, Muller-Werkmeister H, Eberl K, Bredenbeck J, Schwalbe H, Silvers R. The Impact of Azidohomoalanine Incorporation on Protein Structure and Ligand Binding. Chembiochem. 2017 Sep 26. doi: 10.1002/cbic.201700437. PMID:28950050 doi:http://dx.doi.org/10.1002/cbic.201700437
