|
Superimpose homologs
|
POSA server: http://fatcat.burnham.org/POSA/
|
|
ID shared invariant subset
|
Usually 10-30% of monomeric forms
|
|
Re-design exposed surface
|
Protein design: Rosetta http://www.rosettacommons.org/
|
|
Clone & Express
|
Maltose Binding Protein (MBP) fusions; TEV cleavage
|
|
Characterize
|
Rate accelerations, substrate specificities
|
|
Authenticate
|
Single turnover active-site titration, Steady-state kinetics, genetic mutation and manipulation
|
|
Evolutionary recapitulation
|
Multi-scale modular de- and reconstruction
|
