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Biomol NMR Assign
2016 Oct 01;102:407-11. doi: 10.1007/s12104-016-9677-8.
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(1)H, (13)C and (15)N resonance assignments of the Cdc42-binding domain of TOCA1.
Watson JR, Nietlispach D, Owen D, Mott HR.
???displayArticle.abstract??? TOCA1 is a downstream effector protein of the small GTPase, Cdc42. It is a multi-domain protein that includes a membrane binding F-BAR domain, a homology region 1 (HR1) domain, which binds selectively to active Cdc42 and an SH3 domain. TOCA1 is involved in the regulation of actin dynamics in processes such as endocytosis, filopodia formation, neurite elongation, cell motility and invasion. Structural insight into the interaction between TOCA1 and Cdc42 will contribute to our understanding of the role of TOCA1 in actin dynamics. The (1)H, (15)N and (13)C NMR backbone and sidechain resonance assignment of the HR1 domain (12 kDa) presented here provides the foundation for structural studies of the domain and its interactions.
Fig. 1. A 15N-HSQC recorded on 13C, 15N-labelled TOCA1 HR1 domain in 20 mM sodium phosphate pH 7.5, 150 mM NaCl, 5 mM DTT, 5 mM MgCl2, recorded at 500 MHz at 25 °C
Fig. 2. Secondary structure summary of the TOCA1 HR1 domain. Black bars indicate NOEs for each of the atoms listed. The height of the bar represents the strength of the NOE. The atoms listed depict the following NOEs: dαN (dαδ) = CαH of residue i and the NH of residue i + 1, or CδH of i + 1 in the case of Prolines; dNN(dNδ,dδN) = NH of residue i and the NH of residue i + 1, or CδH in the case of Prolines; dβN (dβδ) = CβH of residue i and either the NH or CδH (Prolines) of residue i + 1. For the last four rows, dxy (i,i + z) is used, where x and y denote the atoms in residues i and i + z respectively. The Chemical Shift Index value (CSI) is calculated from all of the backbone atom chemical shifts. A value of +1 indicates a β-strand and a value of −1 indicates an α-helix. The secondary structure was predicted from the chemical shifts using TALOS-N (Shen and Bax 2013) and is depicted as a cartoon below the CSI. This figure was generated in CCPN ANALYSIS (Vranken et al. 2005)
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