Expansion of the genetic code enables design of a novel "gold" class of green fluorescent proteins.
Much effort has been dedicated to the design of significantly red shifted variants of the green fluorescent protein (GFP) from Aequoria victora (av). These approaches have been based on classical engineering with the 20 canonical amino acids. We report here an expansion of these efforts by incorporation of an amino substituted variant of tryptophan into the "cyan" GFP mutant, which turned it into a "gold" variant. This variant possesses a red shift in emission unprecedented for any avFP, similar to "red" FPs, but with enhanced stability and a very low aggregation tendency. An increasing number of non-natural amino acids are available for chromophore redesign (by engineering of the genetic code) and enable new general strategies to generate novel classes of tailor-made GFP proteins.
Max-Planck-Institut für Biochemie, Am Klopferspitz 18A, D-82152 Martinsried, Germany.
Jae Hyun
JH
Bae
Marina
M
Rubini
Gregor
G
Jung
Georg
G
Wiegand
Markus H J
MH
Seifert
M Kamran
MK
Azim
Jeong Sun
JS
Kim
Andreas
A
Zumbusch
Tad A
TA
Holak
Luis
L
Moroder
Robert
R
Huber
Nediljko
N
Budisa
D000818:Animals; D015195:Drug Design; D004355:Drug Stability; D005815:Genetic Code; D049452:Green Fluorescent Proteins; D066298:In Vitro Techniques; D008164:Luminescent Proteins / Q000737:chemistry* / Q000235:genetics*; D008958:Models, Molecular; D011487:Protein Conformation; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D013050:Spectrometry, Fluorescence; D055672:Static Electricity; D013816:Thermodynamics
D008164:Luminescent Proteins; D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins