Fig. 3
From: Automatic structure-based NMR methyl resonance assignment in large proteins
MethylFLYA performance with reduced data input. a Geminal methyl groups of Leu/Val residues can be linked with a short-mixing time NOESY experiment on an exclusively double methyl-labeled ([13δ1/13δ2]-Leu, [13γ1/13γ2]-Val) protein sample. In the NOESY plane of each Leu/Val methyl resonance, a strong signal from its geminal methyl resonance is observed (right). b Different possibilities for treating Leu/Val methyl resonances in MethylFLYA. Top, left: differentiation of the Leu- and Val-type of methyl resonances and known connectivity between geminal Leu or Val methyl groups. Bottom, left: no differentiation between methyl resonances of the Leu- or Val-type, but known connectivity between geminal methyl groups. Top, right: no differentiation between methyl resonances of the Leu/Val-type, nor knowledge of the geminal methyl connectivity. Bottom, right: stereospecific labelling of Leu/Val-methyl groups (pro-R or pro-S), or double labelling (both pro-R and pro-S). c MethylFLYA results with reduced data input. The percentage of assigned methyl groups given knowledge of all methyl amino acid types and the geminal Leu/Val-methyl connectivity is shown in black (all data). In dark gray (L, V = LV), knowledge of Ile- and Ala-methyl resonance types is assumed, but there is no discrimination between Leu or Val methyl resonance types. The geminal (Leu/Val) methyl resonances are connected. In light gray (2L, 2V), knowledge of all amino-acid types is assumed, but the geminal pairing of Leu/Val resonances is omitted. In silver (L, V = LV, 2LV), neither the amino-acid type nor the geminal pairing is known for Leu/Val methyl resonances. Knowledge of the amino acid types of other resonances (Ala, Ile) is still assumed
