1. The effect of amino acid substitution in the kinase inhibitory region of JAB. (A) Unchanged amino acids are shown as dots, substituted amino acids by a one letter code.
The effect of amino acid substitution in the kinase inhibitory region of JAB. (A) Unchanged amino acids are shown as dots, substituted amino acids by a one letter code. All substitutions except Y65F were introduced into the dN51 backbone. The Y65F mutation was introduced into full‐length JAB. All mutants contain an N‐terminal Myc‐epitope tag. (B) EPO‐dependent reporter gene assay was carried out in the presence of the indicated substitution mutants in 293 cells. Data normalized with the β‐galactosidase activity from duplicate experiments are shown. (C) Binding of mutated JAB to JH1. Upper panel: each mutated JAB was transiently expressed in 293 cells and subjected to pY1007 phosphopeptide‐binding assay (pY1007 Bound). Middle and lower panels: GST–JH1 and each JAB mutant were co‐expressed in 293 cells and precipitated with GSH–Sepharose. Myc‐JAB was detected by immunoblotting with anti‐Myc in the total cell lysate (TCL) or GSH‐bound materials (JH1 Bound).
Hideo Yasukawa et al. EMBO J. 1999;18:
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