Caged phosphoproteins.
We present the chemical and biological synthesis of caged phosphoproteins using the in vitro nonsense codon suppression methodology. Specifically, phosphoamino acid analogues of serine, threonine, and tyrosine with a single photocleavable o-nitrophenylethyl caging group were synthesized as the amino acyl tRNA adducts for insertion into full-length proteins. For this purpose, a novel phosphitylating agent was developed. The successful incorporation of these bulky and charged amino acids into the alpha-subunit of the nicotinic acetyl choline receptor (nAChR) and the vasodilator-stimulated phosphoprotein (VASP) using an in vitro translation system is reported.
Rothman
Deborah M
DM
Department of Chemistry, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA.
Petersson
E James
EJ
Vázquez
M Eugenio
ME
Brandt
Gabriel S
GS
Dougherty
Dennis A
DA
Imperiali
Barbara
B
D017868:Cyclic AMP-Dependent Protein Kinases / Q000737:chemistry / Q000378:metabolism; D010750:Phosphoproteins / Q000096:biosynthesis* / Q000138:chemical synthesis* / Q000737:chemistry; D012345:RNA, Transfer, Amino Acid-Specific / Q000737:chemistry / Q000378:metabolism; D012694:Serine / Q000031:analogs & derivatives* / Q000378:metabolism; D013912:Threonine / Q000031:analogs & derivatives* / Q000378:metabolism
D010750:Phosphoproteins; D012345:RNA, Transfer, Amino Acid-Specific; D013912:Threonine; D012694:Serine; D017868:Cyclic AMP-Dependent Protein Kinases