- Macro-seeding is a common crystallographic technique used to
improve crystal size. Transfer a crystal into a fresh drop con-
taining protein/compounds and reservoir solution. Reduce
the concentration of the precipitant (PEG 3350) in the reser-
voir solution until no new crystals form (typically between 3%
and 5% PEG 3350), but high enough that crystal growth can
be observed over 7–10 days. - Depending on the chemistry of the compounds binding to the
γ-subunit, the RIM may or may not interact. In the case of C2,
we would recommend using the AMPKα1/α2 RIM chimera
for crystallization trials. The advantage of using the AMP/C2
combined approach withα 2 β 1 γ1 is the binding of the two
molecules can be directly compared within the one crystal
structure.
Acknowledgments
These authors are supported by grants from the National Health
and Medical Research Council, the Australian Research Council,
the Victorian Government Operational Infrastructure Support
Scheme, and the Jack Brockhoff Foundation (JBF-4206, 2016),
and C.G.L. is an E.H. Flack Research fellow. B.E.K. and J.S.O. are
NHMRC and ARC Research fellows, respectively.References
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