Section 61
Chapter 60,284

Structural basis for intramolecular interaction of post-translationally modified H-Ras•GTP prepared by protein ligation

Ke, H.; Matsumoto, S.; Murashima, Y.; Taniguchi-Tamura, H.; Miyamoto, R.; Yoshikawa, Y.; Tsuda, C.; Kumasaka, T.; Mizohata, E.; Edamatsu, H.; Kataoka, T.

Febs Letters 591(16): 2470-2481


ISSN/ISBN: 0014-5793
PMID: 28730604
DOI: 10.1002/1873-3468.12759
Accession: 060283722

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Ras undergoes post-translational modifications including farnesylation, proteolysis, and carboxymethylation at the C terminus, which are necessary for membrane recruitment and effector recognition. Full activation of c-Raf-1 requires cooperative interaction of the farnesylated C terminus and the activator region of Ras with its cysteine-rich domain (CRD). However, the molecular basis for this interaction remains unclear because of difficulties in preparing modified Ras in amounts sufficient for structural studies. Here, we use Sortase A-catalyzed protein ligation to prepare modified Ras in sufficient amounts for NMR and X-ray crystallographic analyses. The results show that the farnesylated C terminus establishes an intramolecular interaction with the catalytic domain and brings the farnesyl moiety to the proximity of the activator region, which may be responsible for their cooperative recognition of c-Raf-1-CRD.

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