期刊
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
卷 55, 期 28, 页码 8129-8133出版社
WILEY-V C H VERLAG GMBH
DOI: 10.1002/anie.201601299
关键词
C-terminal 1,2-aminothiol; native chemical ligation; N-terminal thioester; protein-protein interactions; Ras homodimer
资金
- Deutsche Forschungsgemeinschaft, DFG [SPP 1623, SFB 642]
- European Research Council, ERC (ChemBioAP)
- Kekule Mobility Fellowship of the Chemical Industry Fund (FCI)
- German Federal Ministry for Education and Research (Medizinische Chemie, TU Dortmund University) [BMBF 1316053]
Weak transient protein-protein interactions (PPIs) play an essential role in cellular dynamics. However, it is challenging to obtain weak protein complexes owing to their short lifetime. Herein we present a general and facile method for trapping weak PPIs in an unbiased manner using proximity-induced ligations. To expand the chemical ligation spectrum, we developed novel N2N (N-terminus to N-terminus) and C2C (C-terminus to C-terminus) ligation approaches. By using N2C (N-terminus to C-terminus), N2N, and C2C ligations in one pot, the interacting proteins were linked. The weak Ypt1: GDI interaction drove C2C ligation with t(1/2) of 4.8 min and near quantitative conversion. The Ypt1-GDI conjugate revealed that binding of Ypt1 G-domain causes opening of the lipid-binding site of GDI, which can accommodate one prenyl group, giving insights into Rab membrane recycling. Moreover, we used this strategy to trap the KRas homodimer, which plays an important role in Ras signaling.
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