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