title: "Turn-on" protein fluorescence: in situ formation of cyanine dyes.
abstract: Protein reengineering of cellular retinoic acid binding protein II (CRABPII) has yielded a genetically addressable system, capable of binding a profluorophoric chromophore that results in fluorescent protein/chromophore complexes. These complexes exhibit far-red emission, with high quantum efficiencies and brightness and also exhibit excellent pH stability spanning the range of 2-11. In the course of this study, it became evident that single mutations of L121E and R59W were most effective in improving the fluorescent characteristics of CRABPII mutants as well as the kinetics of complex formation. The readily crystallizable nature of these proteins was invaluable to provide clues for the observed spectroscopic behavior that results from single mutation of key residues.
authors.affiliation: Department of Chemistry, Michigan State University , East Lansing, Michigan 48824, United States.
authors.forename: Ipek
authors.initials: I
authors.lastname: Yapici
authors.forename: Kin Sing Stephen
authors.initials: KS
authors.lastname: Lee
authors.forename: Tetyana
authors.initials: T
authors.lastname: Berbasova
authors.forename: Meisam
authors.initials: M
authors.lastname: Nosrati
authors.forename: Xiaofei
authors.initials: X
authors.lastname: Jia
authors.forename: Chrysoula
authors.initials: C
authors.lastname: Vasileiou
authors.forename: Wenjing
authors.initials: W
authors.lastname: Wang
authors.forename: Elizabeth M
authors.initials: EM
authors.lastname: Santos
authors.forename: James H
authors.initials: JH
authors.lastname: Geiger
authors.forename: Babak
authors.initials: B
authors.lastname: Borhan
mesh_terms: D002232:Carbocyanines / Q000138:chemical synthesis / Q000737:chemistry*; D004396:Coloring Agents / Q000138:chemical synthesis / Q000737:chemistry*; D005453:Fluorescence; D006863:Hydrogen-Ion Concentration; D008958:Models, Molecular; D015394:Molecular Structure; D018168:Receptors, Retinoic Acid / Q000737:chemistry* / Q000235:genetics
chemical_list: D002232:Carbocyanines; D004396:Coloring Agents; D018168:Receptors, Retinoic Acid; C082928:retinoic acid binding protein II, cellular