A flexible codon in genomically recoded Escherichia coli permits programmable protein phosphorylation
Biochemical investigation of protein phosphorylation events is limited by inefficient production of the phosphorylated and non-phosphorylated forms of full-length proteins. Here using a genomically recoded strain of E. coli with a flexible UAG codon we produce site-specific serine- or phosphoserine-containing proteins, with purities approaching 90%, from a single recombinant DNA. Specifically, we synthesize human MEK1 kinase with two serines or two phosphoserines, from one DNA template, and demonstrate programmable kinase activity. Programmable protein phosphorylation is poised to help reveal the structural and functional information encoded in the phosphoproteome.
| Medienart: |
E-Artikel |
|---|
| Erscheinungsjahr: |
2015 |
|---|---|
| Erschienen: |
2015 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:6 |
|---|---|
| Enthalten in: |
Nature communications - 6(2015) vom: 09. Sept., Seite 8130 |
| Sprache: |
Englisch |
|---|
| Beteiligte Personen: |
Pirman, Natasha L [VerfasserIn] |
|---|
| Links: |
|---|
| Anmerkungen: |
Date Completed 20.04.2016 Date Revised 23.04.2019 published: Electronic Citation Status MEDLINE |
|---|
| doi: |
10.1038/ncomms9130 |
|---|
| funding: |
|
|---|---|
| Förderinstitution / Projekttitel: |
|
| PPN (Katalog-ID): |
NLM252564812 |
|---|
| LEADER | 01000naa a22002652 4500 | ||
|---|---|---|---|
| 001 | NLM252564812 | ||
| 003 | DE-627 | ||
| 005 | 20231224164358.0 | ||
| 007 | cr uuu---uuuuu | ||
| 008 | 231224s2015 xx |||||o 00| ||eng c | ||
| 024 | 7 | |a 10.1038/ncomms9130 |2 doi | |
| 028 | 5 | 2 | |a pubmed24n0841.xml |
| 035 | |a (DE-627)NLM252564812 | ||
| 035 | |a (NLM)26350500 | ||
| 040 | |a DE-627 |b ger |c DE-627 |e rakwb | ||
| 041 | |a eng | ||
| 100 | 1 | |a Pirman, Natasha L |e verfasserin |4 aut | |
| 245 | 1 | 2 | |a A flexible codon in genomically recoded Escherichia coli permits programmable protein phosphorylation |
| 264 | 1 | |c 2015 | |
| 336 | |a Text |b txt |2 rdacontent | ||
| 337 | |a ƒaComputermedien |b c |2 rdamedia | ||
| 338 | |a ƒa Online-Ressource |b cr |2 rdacarrier | ||
| 500 | |a Date Completed 20.04.2016 | ||
| 500 | |a Date Revised 23.04.2019 | ||
| 500 | |a published: Electronic | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a Biochemical investigation of protein phosphorylation events is limited by inefficient production of the phosphorylated and non-phosphorylated forms of full-length proteins. Here using a genomically recoded strain of E. coli with a flexible UAG codon we produce site-specific serine- or phosphoserine-containing proteins, with purities approaching 90%, from a single recombinant DNA. Specifically, we synthesize human MEK1 kinase with two serines or two phosphoserines, from one DNA template, and demonstrate programmable kinase activity. Programmable protein phosphorylation is poised to help reveal the structural and functional information encoded in the phosphoproteome | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, N.I.H., Extramural | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
| 650 | 4 | |a Research Support, U.S. Gov't, Non-P.H.S. | |
| 650 | 7 | |a Codon, Terminator |2 NLM | |
| 650 | 7 | |a Green Fluorescent Proteins |2 NLM | |
| 650 | 7 | |a 147336-22-9 |2 NLM | |
| 650 | 7 | |a Phosphoserine |2 NLM | |
| 650 | 7 | |a 17885-08-4 |2 NLM | |
| 650 | 7 | |a Serine |2 NLM | |
| 650 | 7 | |a 452VLY9402 |2 NLM | |
| 650 | 7 | |a MAP Kinase Kinase 1 |2 NLM | |
| 650 | 7 | |a EC 2.7.12.2 |2 NLM | |
| 650 | 7 | |a MAP2K1 protein, human |2 NLM | |
| 650 | 7 | |a EC 2.7.12.2 |2 NLM | |
| 700 | 1 | |a Barber, Karl W |e verfasserin |4 aut | |
| 700 | 1 | |a Aerni, Hans R |e verfasserin |4 aut | |
| 700 | 1 | |a Ma, Natalie J |e verfasserin |4 aut | |
| 700 | 1 | |a Haimovich, Adrian D |e verfasserin |4 aut | |
| 700 | 1 | |a Rogulina, Svetlana |e verfasserin |4 aut | |
| 700 | 1 | |a Isaacs, Farren J |e verfasserin |4 aut | |
| 700 | 1 | |a Rinehart, Jesse |e verfasserin |4 aut | |
| 773 | 0 | 8 | |i Enthalten in |t Nature communications |d 2010 |g 6(2015) vom: 09. Sept., Seite 8130 |w (DE-627)NLM199274525 |x 2041-1723 |7 nnns |
| 773 | 1 | 8 | |g volume:6 |g year:2015 |g day:09 |g month:09 |g pages:8130 |
| 856 | 4 | 0 | |u http://dx.doi.org/10.1038/ncomms9130 |3 Volltext |
| 912 | |a GBV_USEFLAG_A | ||
| 912 | |a GBV_NLM | ||
| 951 | |a AR | ||
| 952 | |d 6 |j 2015 |b 09 |c 09 |h 8130 | ||