Caging proteins through unnatural amino acid mutagenesis.
The caging of specific residues of proteins is a powerful tool. This discussion attempts to alert the reader to the considerations that must be made in preparing and analyzing a caged protein through nonsense suppression. Although the suppression methodology is conceptually straightforward, it not possible to provide a failsafe "cook book" method for using caged unnaturals. We have emphasized the preparation of caged receptors expressed in Xenopus oocytes, but these approaches can clearly be adapted to many other systems.
Petersson
E James
EJ
Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena 91125, USA.
Brandt
Gabriel S
GS
Zacharias
Niki M
NM
Dougherty
Dennis A
DA
Lester
Henry A
HA
D000596:Amino Acids / Q000737:chemistry* / Q000235:genetics; D000818:Animals; D018388:Codon, Terminator; D016296:Mutagenesis; D012333:RNA, Messenger / Q000235:genetics; D014158:Transcription, Genetic; D014981:Xenopus
D000596:Amino Acids; D018388:Codon, Terminator; D012333:RNA, Messenger