TY - JOUR
T1 - A rationally designed orthogonal synthetase for genetically encoded fluorescent amino acids
AU - Steinberg, Ximena
AU - Galpin, Jason
AU - Nasir, Gibran
AU - Sepúlveda, Romina V.
AU - Ladron de Guevara, Ernesto
AU - Gonzalez-Nilo, Fernando
AU - Islas, Leon D.
AU - Ahern, Christopher A.
AU - Brauchi, Sebastian E.
N1 - Funding Information:
This work was supported by DRI-CONICYT/CONACYT grant to S. Brauchi and L.D.Islas (Grant number PCCI12023 ), Iniciativa Cientifica Milenio (ANID , Chilean Government), and the National Institutes of Health (U.S. Government). F. Gonzalez-Nilo was supported by FONDECYT (grant number 1170733 ) and Grand Laureate. L. Islas was supported by DGAPA-PAPIIT-UNAM (grant number IN209515 and IN203318 ). C. Ahern was supported by National Institutes of Health (NIGMS grant numbers GM106569 and GM087519 ). S. Brauchi was supported by FONDECYT (grant number 1191868 ) and Anillo Cientifico (grant number ACT-1401).
Funding Information:
X. Steinberg was a MECESUP and CONICYT fellow; R.V. Sepulveda was a CONICYT fellow. C.A.Ahern is a member of the Membrane Protein Structural Dynamics Consortium, which is funded by NIH. S.E.Brauchi is a member of the Millennium Nucleus of Ion-Channel Associated Diseases (MiNICAD), which is funded by ANID.
Publisher Copyright:
© 2020
Copyright:
Copyright 2020 Elsevier B.V., All rights reserved.
PY - 2020/10
Y1 - 2020/10
N2 - The incorporation of non-canonical amino acids into proteins has emerged as a promising strategy to manipulate and study protein structure-function relationships with superior precision in vitro and in vivo. To date, fluorescent non-canonical amino acids (f-ncAA) have been successfully incorporated in proteins expressed in bacterial systems, Xenopus oocytes, and HEK-293T cells. Here, we describe the rational generation of a novel orthogonal aminoacyl-tRNA synthetase based on the E. coli tyrosine synthetase that is capable of encoding the f-ncAA tyr-coumarin in HEK-293T cells.
AB - The incorporation of non-canonical amino acids into proteins has emerged as a promising strategy to manipulate and study protein structure-function relationships with superior precision in vitro and in vivo. To date, fluorescent non-canonical amino acids (f-ncAA) have been successfully incorporated in proteins expressed in bacterial systems, Xenopus oocytes, and HEK-293T cells. Here, we describe the rational generation of a novel orthogonal aminoacyl-tRNA synthetase based on the E. coli tyrosine synthetase that is capable of encoding the f-ncAA tyr-coumarin in HEK-293T cells.
KW - Aminoacyl-tRNA synthetase
KW - Biochemistry
KW - Bioinformatics
KW - Bioorganic chemistry
KW - Coumarin
KW - Fluorescence
KW - Molecular biology
KW - Proteins
KW - Unnatural amino acids
UR - http://www.scopus.com/inward/record.url?scp=85092260461&partnerID=8YFLogxK
U2 - 10.1016/j.heliyon.2020.e05140
DO - 10.1016/j.heliyon.2020.e05140
M3 - Article
AN - SCOPUS:85092260461
SN - 2405-8440
VL - 6
JO - Heliyon
JF - Heliyon
IS - 10
M1 - e05140
ER -