Generating enzyme and radical-mediated bisubstrates as tools for investigating Gcn5-related N-acetyltransferases
© 2017 Federation of European Biochemical Societies..
Gcn5-related N-acetyltransferases (GNATs) are found in all kingdoms of life and catalyze important acyl transfer reactions in diverse cellular processes. While many 3D structures of GNATs have been determined, most do not contain acceptor substrates in their active sites. To expand upon existing crystallographic strategies for improving acceptor-bound GNAT structures, we synthesized peptide substrate analogs and reacted them with CoA in PA4794 protein crystals. We found two separate mechanisms for bisubstrate formation: (a) a novel X-ray induced radical-mediated alkylation of CoA with an alkene peptide and (b) direct alkylation of CoA with a halogenated peptide. Our approach is widely applicable across the GNAT superfamily and can be used to improve the success rate of obtaining liganded structures of other acyltransferases.
| Medienart: |
E-Artikel |
|---|
| Erscheinungsjahr: |
2017 |
|---|---|
| Erschienen: |
2017 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:591 |
|---|---|
| Enthalten in: |
FEBS letters - 591(2017), 15 vom: 19. Aug., Seite 2348-2361 |
| Sprache: |
Englisch |
|---|
| Beteiligte Personen: |
Reidl, Cory [VerfasserIn] |
|---|
| Links: |
|---|
| Anmerkungen: |
Date Completed 10.10.2017 Date Revised 06.01.2023 published: Print-Electronic Citation Status MEDLINE |
|---|
| doi: |
10.1002/1873-3468.12753 |
|---|
| funding: |
|
|---|---|
| Förderinstitution / Projekttitel: |
|
| PPN (Katalog-ID): |
NLM273794140 |
|---|
| LEADER | 01000naa a22002652 4500 | ||
|---|---|---|---|
| 001 | NLM273794140 | ||
| 003 | DE-627 | ||
| 005 | 20231225002033.0 | ||
| 007 | cr uuu---uuuuu | ||
| 008 | 231225s2017 xx |||||o 00| ||eng c | ||
| 024 | 7 | |a 10.1002/1873-3468.12753 |2 doi | |
| 028 | 5 | 2 | |a pubmed24n0912.xml |
| 035 | |a (DE-627)NLM273794140 | ||
| 035 | |a (NLM)28703494 | ||
| 040 | |a DE-627 |b ger |c DE-627 |e rakwb | ||
| 041 | |a eng | ||
| 100 | 1 | |a Reidl, Cory |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Generating enzyme and radical-mediated bisubstrates as tools for investigating Gcn5-related N-acetyltransferases |
| 264 | 1 | |c 2017 | |
| 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 10.10.2017 | ||
| 500 | |a Date Revised 06.01.2023 | ||
| 500 | |a published: Print-Electronic | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a © 2017 Federation of European Biochemical Societies. | ||
| 520 | |a Gcn5-related N-acetyltransferases (GNATs) are found in all kingdoms of life and catalyze important acyl transfer reactions in diverse cellular processes. While many 3D structures of GNATs have been determined, most do not contain acceptor substrates in their active sites. To expand upon existing crystallographic strategies for improving acceptor-bound GNAT structures, we synthesized peptide substrate analogs and reacted them with CoA in PA4794 protein crystals. We found two separate mechanisms for bisubstrate formation: (a) a novel X-ray induced radical-mediated alkylation of CoA with an alkene peptide and (b) direct alkylation of CoA with a halogenated peptide. Our approach is widely applicable across the GNAT superfamily and can be used to improve the success rate of obtaining liganded structures of other acyltransferases | ||
| 650 | 4 | |a Letter | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
| 650 | 4 | |a Research Support, N.I.H., Extramural | |
| 650 | 4 | |a Research Support, U.S. Gov't, Non-P.H.S. | |
| 650 | 4 | |a Gcn5-related N-acetyltransferase | |
| 650 | 4 | |a product-based transition-state modeling | |
| 650 | 4 | |a radical-mediated bisubstrate | |
| 650 | 7 | |a Dipeptides |2 NLM | |
| 650 | 7 | |a Acetyltransferases |2 NLM | |
| 650 | 7 | |a EC 2.3.1.- |2 NLM | |
| 650 | 7 | |a Coenzyme A |2 NLM | |
| 650 | 7 | |a SAA04E81UX |2 NLM | |
| 700 | 1 | |a Majorek, Karolina A |e verfasserin |4 aut | |
| 700 | 1 | |a Dang, Joseph |e verfasserin |4 aut | |
| 700 | 1 | |a Tran, David |e verfasserin |4 aut | |
| 700 | 1 | |a Jew, Kristen |e verfasserin |4 aut | |
| 700 | 1 | |a Law, Melissa |e verfasserin |4 aut | |
| 700 | 1 | |a Payne, Yasmine |e verfasserin |4 aut | |
| 700 | 1 | |a Minor, Wladek |e verfasserin |4 aut | |
| 700 | 1 | |a Becker, Daniel P |e verfasserin |4 aut | |
| 700 | 1 | |a Kuhn, Misty L |e verfasserin |4 aut | |
| 773 | 0 | 8 | |i Enthalten in |t FEBS letters |d 1968 |g 591(2017), 15 vom: 19. Aug., Seite 2348-2361 |w (DE-627)NLM000016691 |x 1873-3468 |7 nnns |
| 773 | 1 | 8 | |g volume:591 |g year:2017 |g number:15 |g day:19 |g month:08 |g pages:2348-2361 |
| 856 | 4 | 0 | |u http://dx.doi.org/10.1002/1873-3468.12753 |3 Volltext |
| 912 | |a GBV_USEFLAG_A | ||
| 912 | |a GBV_NLM | ||
| 951 | |a AR | ||
| 952 | |d 591 |j 2017 |e 15 |b 19 |c 08 |h 2348-2361 | ||