A six-dimensional alpha proton detection-based APSY experiment for backbone assignment of intrinsically disordered proteins

Liquid–liquid phase separation of the microtubule-binding repeats of the Alzheimer-related protein Tau

“…a Two-dimensional 1 H- 15 N HSQC spectra of the repeat domain of Tau (100 μM of K18 in 50 mM sodium phosphate, pH 8.8, and 0.5 mM TCEP) in the monomeric dispersed ( blue ; 5 °C) and liquid demixed ( red ; 37 °C) state. Due to increased solvent exchange, many 1 H- 15 N cross-peaks of K18 are attenuated beyond detection at 37 °C 88 . In addition, an overall shift of the backbone resonances was observed, which is caused by the temperature dependence of NMR chemical shifts 52 .…”

Focusing on “the important” through targeted NMR experiments: an example of selective13C–12C bond detection in complex mixtures

“…7 On extrapolation to 7D NMR, which has recently been demonstrated on disordered proteins via sparse sampling approaches, the peak capacity can potentially reach on the order of 10 18 . [8][9][10] Of course, in most environmental and biological samples, the limiting factor is a lack of sensitivity when dispersing signals into a larger number of higher dimensions. However, as NMR sensitivity continues to increase and sparse sampling approaches improve, 11 NMR experiments of high dimensionality become more feasible, and will therefore likely become central to the next generation of complex mixture research.…”

Six- and seven-dimensional experiments by combination of sparse random sampling and projection spectroscopy dedicated for backbone resonance assignment of intrinsically disordered proteins

“…To date, several high dimensional strategies have been proposed for backbone resonance assignment of IDPs (Reddy and Hosur 2014 ; Goradia et al 2015 ; Yoshimura et al 2015 ), including direct 13 C detection (Bermel et al 2006 , 2009 ; Nováček et al 2011 ; Pantoja-Uceda and Santoro 2013 , 2014 ) and HA detection (Mäntylahti et al 2010 , 2011 ; Yao et al 2014 ) which allow for revealing of proline residues. The number of dimensions, i.e.…”

“…The number of dimensions, i.e. number of simultaneously correlated frequencies, could be increased by employing automated projection spectroscopy (APSY) (Hiller et al 2007 ; Narayanan et al 2010 ; Yao et al 2014 ), or the non-uniformly sampled (NUS) high-dimensional experiments (Kazimierczuk et al 2010 ; Zawadzka-Kazimierczuk et al 2012 ).…”