Journal
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
Volume 61, Issue 1, Pages -Publisher
WILEY-V C H VERLAG GMBH
DOI: 10.1002/anie.202108361
Keywords
IDP datasets; NMR H-1(alpha)-detection; p53; phosphorylation; proline cis/trans isomerization
Categories
Funding
- National Research, Development and Innovation Office [TKP2020-IKA-05, 2018-1.2.1-NKP-2018-00005]
- NKFI [K124900, K137940, K119359, EFOP-3.6.3-VEKOP-16-2017-00009]
- Foundation for Hungarian Peptide and Protein Research
- HGF program Information [43.35.02]
- Virtmat
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This study introduces two new NMR experiments to identify proline cis/trans isomers in proteins, detecting minor conformers present in 4-15%, and analyzes the impact of CK2 phosphorylation on the cis/trans-proline equilibrium. Statistical analysis indicates that the amino acid type also affects the distribution of cis/trans-proline.
It is important to identify proline cis/trans isomers that appear in several regulatory mechanisms of proteins, and to characterize minor species that are present due to the conformational heterogeneity in intrinsically disordered proteins (IDPs). To obtain residue level information on these mobile systems we introduce two H-1(alpha)-detected, proline selective, real-time homodecoupled NMR experiments and analyze the proline abundant transactivation domain of p53. The measurements are sensitive enough to identify minor conformers present in 4-15 % amounts; moreover, we show the consequences of CK2 phosphorylation on the cis/trans-proline equilibrium. Using our results and available literature data we perform a statistical analysis on how the amino acid type effects the cis/trans-proline distribution. The methods are applicable under physiological conditions, they can contribute to find key proline isomers in proteins, and statistical analysis results may help in amino acid sequence optimization for biotechnological purposes.
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