Design of Light-Controlled Protein Conformations and Functions.
In recent years, interest in controlling protein function with light has increased. Light offers a number of unique advantages over other methods, including spatial and temporal control and high selectivity. Here, we describe a general protocol for engineering a protein to be controllable with light via reaction with an exogenously introduced photoisomerizable small molecule and illustrate our protocol with two examples from the literature: the engineering of the calcium affinity of the cell-cell adhesion protein cadherin, which is an example of a protein that switches from a native to a disrupted state (Ritterson et al. J Am Chem Soc (2013) 135:12516-12519), and the engineering of the opening and closing of the chaperonin Mm-cpn, an example of a switch between two functional states (Hoersch et al.: Nat Nanotechn (2013) 8:928-932). This protocol guides the user from considering which proteins may be most amenable to this type of engineering, to considerations of how and where to make the desired changes, to the assays required to test for functionality.
California Institute for Quantitative Biomedical Research and Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, 1700 4th Street, Byers Hall 308 E, San Francisco, CA, 94158, USA.
Ryan S
RS
Ritterson
California Institute for Quantitative Biomedical Research and Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, 1700 4th Street, Byers Hall 308 E, San Francisco, CA, 94158, USA.|Fachbereich Physik, Freie Universität Berlin, Arnimallee 14, 14195, Berlin, Germany.
Daniel
D
Hoersch
Graduate Program in Bioinformatics, California Institute for Quantitative Biomedical Research, and Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, 1700 4th Street, Byers Hall 308 E, San Francisco, CA, 94158, USA.
Kyle A
KA
Barlow
California Institute for Quantitative Biomedical Research and Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, 1700 4th Street, Byers Hall 308 E, San Francisco, CA, 94158, USA. kortemme@cgl.ucsf.edu.
Tanja
T
Kortemme
D004591:Electrophoresis, Polyacrylamide Gel; D008027:Light; D013058:Mass Spectrometry; D009154:Mutation; D011487:Protein Conformation; D013056:Spectrophotometry, Ultraviolet
Computational protein design; Light-modulatable proteins; Photoswitches; Protein engineering