title: Optical control of protein activity by fluorescent protein domains.
abstract: Fluorescent proteins (FPs) are widely used as optical sensors, whereas other light-absorbing domains have been used for optical control of protein localization or activity. Here, we describe light-dependent dissociation and association in a mutant of the photochromic FP Dronpa, and we used it to control protein activities with light. We created a fluorescent light-inducible protein design in which Dronpa domains are fused to both termini of an enzyme domain. In the dark, the Dronpa domains associate and cage the protein, but light induces Dronpa dissociation and activates the protein. This method enabled optical control over guanine nucleotide exchange factor and protease domains without extensive screening. Our findings extend the applications of FPs from exclusively sensing functions to also encompass optogenetic control.
authors.affiliation: Department of Bioengineering, Stanford University, Stanford, CA 94305, USA.
authors.forename: Xin X
authors.initials: XX
authors.lastname: Zhou
authors.forename: Hokyung K
authors.initials: HK
authors.lastname: Chung
authors.forename: Amy J
authors.initials: AJ
authors.lastname: Lam
authors.forename: Michael Z
authors.initials: MZ
authors.lastname: Lin
mesh_terms: D033942:Adaptor Proteins, Vesicular Transport / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D003624:Darkness; D005453:Fluorescence; D006367:HeLa Cells; D006801:Humans; D008027:Light; D008164:Luminescent Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008958:Models, Molecular; D041681:NIH 3T3 Cells; D063347:Native Polyacrylamide Gel Electrophoresis; D062308:Optogenetics; D011487:Protein Conformation; D015202:Protein Engineering; D055503:Protein Multimerization; D017434:Protein Structure, Tertiary; D011554:Pseudopodia / Q000378:metabolism / Q000648:ultrastructure; D011993:Recombinant Fusion Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D012697:Serine Endopeptidases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D017361:Viral Nonstructural Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases
chemical_list: D033942:Adaptor Proteins, Vesicular Transport; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D012697:Serine Endopeptidases; D017361:Viral Nonstructural Proteins; C084422:NS3 protein, hepatitis C virus; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases; C093542:NS4 protein, hepatitis C virus; C115919:intersectin 1; C547377:neptune protein