ncAA Structure (png, jpg, jpeg)
ncAA Foundational Publication Support
Rogerson, D. T., A. Sachdeva, K. Wang, T. Haq, A. Kazlauskaite, S. M. Hancock, N. Huguenin-Dezot, M. M. K. Muqit, A. M. Fry, and R. Bayliss. n.d. “Efficient Genetic Encoding Of Phosphoserine And Its Nonhydrolyzable Analog” 11: 496+.
Beránek, V., C. D. Reinkemeier, M. S. Zhang, A. D. Liang, G. Kym, and J. W. Chin. n.d. “Genetically Encoded Protein Phosphorylation In Mammalian Cells” 25: 1067+.
Zhu, Phillip, Stanislau Stanisheuski, Rachel Franklin, Amber Vogel, Cat Hoang Vesely, Patrick Reardon, Nikolai N Sluchanko, et al. (2023) 2023. “Autonomous Synthesis Of Functional, Permanently Phosphorylated Proteins For Defining The Interactome Of Monomeric 14-3-3Ζ.”. Acs Central Science 9 (4): 816-835. doi:10.1021/acscentsci.3c00191.
ncAA Protocols
Zhu, Phillip, Ryan A Mehl, and Richard B Cooley. (2023) 2023. “Biosynthesis And Genetic Encoding Of Non-Hydrolyzable Phosphoserine Into Recombinant Proteins In Escherichia Coli.”. Bio-Protocol 13 (21): e4861. doi:10.21769/BioProtoc.4861.
ncAA Usage Publications
Stuber, Katrin, Tobias Schneider, Jill Werner, Michael Kovermann, Andreas Marx, and Martin Scheffner. (2021) 2021. “Structural And Functional Consequences Of Nedd8 Phosphorylation.”. Nature Communications 12 (1): 5939. doi:10.1038/s41467-021-26189-9.
Liu, Dong, Yingying Liu, Hua-Zhen Duan, Xinjie Chen, Yanan Wang, Ting Wang, Qing Yu, Yong-Xiang Chen, and Yuan Lu. (2023) 2023. “Customized Synthesis Of Phosphoprotein Bearing Phosphoserine Or Its Nonhydrolyzable Analog.”. Synthetic And Systems Biotechnology 8 (1): 69-78. doi:10.1016/j.synbio.2022.11.004.
ncAA Utility
Used to produce proteins that mimic site-specific phosphorylation at a serine residue - where the phosphate group cannot by hydrolyzed - via genetic code expansion versus other protein phosphorylation strategies.
ncAA Source
Endogenous - Biosynthesized
ncAA Availability
In the most recent paper (Zhu et al. 2023) - nhpSer is biosynthesized via the Frb protein pathway. The "PermaPhos" kit is available on Addgene kit #1000000226.
For mammalian expressions, exogenous nhpSer is added to the media.
For mammalian expressions, exogenous nhpSer is added to the media.
RS/tRNA Pair Usage Information
For incorporating nhpSer in E. coli, the Permaphos system using endogenously synthesized nhpSer and using the SepRS2/Sep-tRNA-v2 pair performs best.
For mammalian expressions, the 2018 system built on the SepRS2/Sep-tRNA(B4) pair gives ~50% fidelity.
For mammalian expressions, the 2018 system built on the SepRS2/Sep-tRNA(B4) pair gives ~50% fidelity.
ncAA Synonyms
nhpSer
nonhydrolyzable phosphoserine
non-hydrolyzable phosphoserine
phosphono-methyl-alanine
2-amino-4-phosphonobutyric acid
2-amino-4-phosphonobutyrate
L-2-amino-4-phosphonobutyric acid
(2S)-2-amino-4-phosphonobutanoic acid
(2S)-2-amino-4-phosphonobutyric acid
DL-AP4
nonhydrolyzable phosphoserine
non-hydrolyzable phosphoserine
phosphono-methyl-alanine
2-amino-4-phosphonobutyric acid
2-amino-4-phosphonobutyrate
L-2-amino-4-phosphonobutyric acid
(2S)-2-amino-4-phosphonobutanoic acid
(2S)-2-amino-4-phosphonobutyric acid
DL-AP4
ChEBI ID
143992
PubChem Link
PDB Code
E7P