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Figure 7.
The C-terminal hydrophobic residues (Ile83 and Leu84) of AscF
are essential for chaperone binding. (Lane 1) Protein molecular
weight marker; (lane 2) coexpression of full-length His-AscF
with AscE and AscG. The formation of the ternary complex is
indicated by copurification of AscE (7.56 kDa). (Lane 3)
Coexpression of the His-AscF^[Delta]81 --87 C-terminal
truncation mutant with AscE and AscG. No ternary complex can be
formed, as indicated by the absence of the band corresponding to
AscE. (Lanes 4 --7) Coexpression of His-AscF I83A_L84A, His-AscF
I83A_I87A, His-AscF L84A_I87A, and His-AscF I83A_L84A_I87A,
respectively, with AscE and AscG. Mutation of both residues
Ile83 and Leu84 are necessary to reduce the chaperone binding of
AscF significantly. Residue Ile87 is also involved in the
interaction but not to the same extent as residues Ile83 and
Leu84.
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