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Structural basis for intramolecular interaction of post‐translationally modified H‐Ras• GTP prepared by protein ligation
Author(s) -
Ke Haoliang,
Matsumoto Shigeyuki,
Murashima Yosuke,
TaniguchiTamura Haruka,
Miyamoto Ryo,
Yoshikawa Yoko,
Tsuda Chiemi,
Kumasaka Takashi,
Mizohata Eiichi,
Edamatsu Hironori,
Kataoka Tohru
Publication year - 2017
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1002/1873-3468.12759
Subject(s) - prenylation , chemistry , moiety , n terminus , activator (genetics) , c terminus , gtp' , gtpase , cysteine , stereochemistry , intramolecular force , biochemistry , microbiology and biotechnology , peptide sequence , biology , receptor , amino acid , enzyme , gene
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 .