5MZ7

Crystal Structure of the third PDZ domain from the synaptic protein PSD-95 with incorporated Azidohomoalanine

5MZ7 の概要

• エントリーDOI: 10.2210/pdb5mz7/pdb
• 分子名称: Disks large homolog 4 (2 entities in total)
• 機能のキーワード: azidohomoalanine, non-standard amino acid, unnatural amino acid, artificial amino acid, psd-95, peptide binding protein
• 由来する生物種: Rattus norvegicus (Norway rat)
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 43211.80

構造登録者

Kudlinzki, D.,Lehner, F.,Witt, K.,Linhard, V.L.,Silvers, R.,Schwalbe, H. (登録日: 2017-01-31, 公開日: 2017-10-04, 最終更新日: 2024-01-17)

主引用文献

Lehner, F.,Kudlinzki, D.,Richter, C.,Muller-Werkmeister, H.M.,Eberl, K.B.,Bredenbeck, J.,Schwalbe, H.,Silvers, R.
Impact of Azidohomoalanine Incorporation on Protein Structure and Ligand Binding.
Chembiochem, 18:2340-2350, 2017

PubMed Abstract: The impact of the incorporation of a non-natural amino acid (NNAA) on 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 through click chemistry. Herein, structural characterisation of the impact of the incorporation of azidohomoalanine (AZH) into the model protein domain PDZ3 is examined by means of NMR spectroscopy and X-ray crystallography. The structure and dynamics of the apo state of AZH-modified PDZ3 remain mostly unperturbed. Furthermore, the binding of two PDZ3 binding peptides are unchanged upon incorporation of AZH. The interface of the AZH-modified PDZ3 and an azulene-linked peptide for vibrational energy transfer studies has been mapped by means of chemical shift perturbations and NOEs between the unlabelled azulene-linked peptide and the isotopically labelled protein. Co-crystallisation and soaking failed for the peptide-bound holo complex. NMR spectroscopy, however, allowed determination of the protein-ligand interface. Although the incorporation of AZH was minimally invasive for PDZ3, structural analysis of NNAA-modified proteins through the methodology presented herein should be performed to ensure structural integrity of the studied target.

PubMed: 28950050
DOI: 10.1002/cbic.201700437

実験手法

X-RAY DIFFRACTION (1.53 Å)