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Dear CCP4bb experts:
I am sorry for the delay in replying you guys.
Thank a lot for your good ideas, including soaking, cross-linking, creating
new fusion constructs and trying to purify the complex by PEG precipitation,
and so on.
For further discussion, I would like to give some detailed information about
my complex.
The Biacore3000 gave a dissociation constant of 3.0e-5 M using 2mM
Hepes(pH7.4), 0.15M NaCl as the running buffer.
I think this constant is equal to a protein concentration of mg/ml level.
One more thing that I want to mention is that the complex would like to
precipitate during concentration or at 4 degree(according to the SDS-Gel),
and the complex pellet dissolved a little better in the buffer containing
0.15M NaCl than that containing 0.05M NaCl.
The MW of the big component is 80KD, and 20KD for the small one. One Asn
residue in the big component is important for the binding. But I do not know
whether it forms ion pair with the small one.
Would everybody like to give me some more advice?
Thanks a million.
Jerry McCully
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