Intermolecular NOEs observed in the IIAMan-IIBMan
complex. Methyl region of 1H(13C-attached,
F3)/13C(F2) planes from a three-dimensional
12C-filtered(F1)/13C-separated(F2)
NOE-HSQC spectrum displaying NOEs from the methyl protons of Met-103 (top
panels), Met-23 (middle panels), and Leu-129 δ1
(bottom panels) of
[12CH3-LV]/[1H-MV]/[2H/12C/14N]-IIAMan
to protons attached to
13Cof[U-15N/13C]-IIBMan.
A, wild-type IIAMan(H10) and IIBMan.
B, IIAMan(H10E) and IIBMan. C,
location of the two clusters of residues located on helices α1 and
α3 of IIBMan that are far apart in the structure but exhibit
NOEs to the methyl group of Met-103 in the wild-type sample. The wild-type
sample (A) displays NOEs arising from a mixture of two complexes
comprising one that is compatible with phosphoryl transfer (productive) and
the other where phosphoryl transfer cannot take place (non-productive); the
complex of the phosphomimetic IIAMan(H10E) with IIBMan
(B) corresponds to a single productive complex.