Chemical shift based editing of CH3 groups in fractionally 13C-labelled proteins using GFT (3, 2)D CT-HCCH-COSY: stereospecific assignments of CH3 groups of Val and Leu residues.
We propose a (3, 2)D CT-HCCH-COSY experiment to rapidly collect the data and provide significant dispersion in the spectral region containing (13)C-(1)H cross peaks of CH(3) groups belonging to Ala, Ile, Leu, Met, Thr and Val residues. This enables one to carry out chemical shift based editing and grouping of all the (13)C-(1)H cross peaks of CH(3) groups belonging to Ala, Ile, Leu, Met, Thr and Val residues in fractionally (10%) (13)C-labelled proteins, which in turn aids in the sequence-specific resonance assignments in general and side-chain resonance assignments in particular, in any given protein. Further, we demonstrate the utility of this experiment for stereospecific assignments of the pro-R and pro-S methyl groups belonging to the Leu and Val residues in fractionally (10%) (13)C-labelled proteins. The proposed experiment opens up a wide range of applications in resonance assignment strategies and structure determination of proteins.
Barnwal
Ravi Pratap
RP
Department of Chemical Sciences, Tata Institute of Fundamental Research, Colaba, Mumbai, India.
Atreya
Hanudatta S
HS
Chary
Kandala V R
KV
D002247:Carbon Isotopes / Q000737:chemistry*; D003459:Crystallins / Q000737:chemistry; D007930:Leucine / Q000737:chemistry*; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D011506:Proteins / Q000737:chemistry*; D014633:Valine / Q000737:chemistry*; D000090803:mu-Crystallins
D002247:Carbon Isotopes; D003459:Crystallins; D011506:Proteins; D000090803:mu-Crystallins; D007930:Leucine; D014633:Valine