一种用于分离模块化合成酶交联结构域的正交纯化策略。
An orthogonal purification strategy for isolating crosslinked domains of modular synthases.
作者信息
Haushalter Robert W, Worthington Andrew S, Hur Gene H, Burkart Michael D
机构信息
Department of Chemistry and Biochemistry, University of California, San Diego, 9500 Gilman Drive, La Jolla, CA 92093-0358, USA.
出版信息
Bioorg Med Chem Lett. 2008 May 15;18(10):3039-42. doi: 10.1016/j.bmcl.2008.01.026. Epub 2008 Jan 11.
Abstract
Chemo-enzymatic methods for covalently crosslinking carrier proteins with partner enzymes within modular synthases hold promise for elucidating and engineering metabolic pathways. Our efforts to crystallize the ACP-KS complexes of fatty acid synthases have been complicated by difficulties in the purification of the crosslinked complex from the other proteins in the reaction. Here we present a solution that employs an orthogonal purification strategy to achieve the quantity and level of purity necessary for further studies of this complex.
摘要
在模块化合成酶中，通过化学酶法将载体蛋白与伴侣酶共价交联的方法，有望用于阐明和改造代谢途径。我们在结晶脂肪酸合成酶的ACP-KS复合物时，由于难以从反应中的其他蛋白质中纯化交联复合物而变得复杂。在此，我们提出一种解决方案，该方案采用正交纯化策略，以获得进一步研究该复合物所需的数量和纯度水平。
相似文献
1
An orthogonal purification strategy for isolating crosslinked domains of modular synthases.
Bioorg Med Chem Lett. 2008 May 15;18(10):3039-42. doi: 10.1016/j.bmcl.2008.01.026. Epub 2008 Jan 11.
2
Structural basis for selectivity in a highly reducing type II polyketide synthase.
Nat Chem Biol. 2020 Jul;16(7):776-782. doi: 10.1038/s41589-020-0530-0. Epub 2020 May 4.
3
The structural organization of substrate loading in iterative polyketide synthases.
Nat Chem Biol. 2018 May;14(5):474-479. doi: 10.1038/s41589-018-0026-3. Epub 2018 Apr 2.
4
Probing the selectivity and protein·protein interactions of a nonreducing fungal polyketide synthase using mechanism-based crosslinkers.
Chem Biol. 2013 Sep 19;20(9):1135-46. doi: 10.1016/j.chembiol.2013.07.012. Epub 2013 Aug 29.
5
Mechanism based protein crosslinking of domains from the 6-deoxyerythronolide B synthase.
Bioorg Med Chem Lett. 2008 May 15;18(10):3034-8. doi: 10.1016/j.bmcl.2008.01.073. Epub 2008 Jan 24.
6
A photocrosslinking assay for reporting protein interactions in polyketide and fatty acid synthases.
Mol Biosyst. 2011 Nov;7(11):3152-6. doi: 10.1039/c1mb05270e. Epub 2011 Sep 14.
7
Engineered Chimeras Unveil Swappable Modular Features of Fatty Acid and Polyketide Synthase Acyl Carrier Proteins.
Biochemistry. 2022 Feb 15;61(4):217-227. doi: 10.1021/acs.biochem.1c00798. Epub 2022 Jan 24.
8
One-Step Transformation of Coenzyme A into Analogues by Transamidation.
Org Lett. 2017 Apr 21;19(8):1950-1953. doi: 10.1021/acs.orglett.7b00291. Epub 2017 Apr 10.
9
An ACP structural switch: conformational differences between the apo and holo forms of the actinorhodin polyketide synthase acyl carrier protein.
Chembiochem. 2008 Oct 13;9(15):2424-32. doi: 10.1002/cbic.200800180.
10
Solution structure and backbone dynamics of the holo form of the frenolicin acyl carrier protein.
Biochemistry. 2003 Apr 29;42(16):4648-57. doi: 10.1021/bi0274120.
引用本文的文献
1
Elucidation of interface interactions between a dehydratase domain and an acyl carrier protein in cremimycin polyketide synthase.
FEBS Lett. 2025 Apr;599(8):1159-1168. doi: 10.1002/1873-3468.15103. Epub 2025 Jan 26.
2
Fluorometric Analysis of Carrier-Protein-Dependent Biosynthesis through a Conformationally Sensitive Solvatochromic Pantetheinamide Probe.
ACS Chem Biol. 2024 Jul 19;19(7):1416-1425. doi: 10.1021/acschembio.4c00169. Epub 2024 Jun 23.
3
Quantitative Characterization of Chain-Flipping of Acyl Carrier Protein of Using Chemical Exchange NMR.
J Am Chem Soc. 2024 Jul 10;146(27):18650-18660. doi: 10.1021/jacs.4c05509. Epub 2024 Jun 14.
4
Cross-Linking of the Nonribosomal Peptide Synthetase Adenylation Domain with a Carrier Protein Using a Pantetheine-Type Probe.
Methods Mol Biol. 2023;2670:207-217. doi: 10.1007/978-1-0716-3214-7_10.
5
Protein-Protein Recognition Involved in the Intermodular Transacylation Reaction in Modular Polyketide Synthase in the Biosynthesis of Vicenistatin.
Chembiochem. 2022 Jul 19;23(14):e202200200. doi: 10.1002/cbic.202200200. Epub 2022 May 16.
6
Complex structure of the acyltransferase VinK and the carrier protein VinL with a pantetheine cross-linking probe.
Acta Crystallogr F Struct Biol Commun. 2021 Sep 1;77(Pt 9):294-302. doi: 10.1107/S2053230X21008761. Epub 2021 Aug 26.
7
PKS-NRPS Enzymology and Structural Biology: Considerations in Protein Production.
Methods Enzymol. 2018;604:45-88. doi: 10.1016/bs.mie.2018.01.035. Epub 2018 Mar 16.
8
Matching Protein Interfaces for Improved Medium-Chain Fatty Acid Production.
ACS Synth Biol. 2018 May 18;7(5):1179-1187. doi: 10.1021/acssynbio.7b00334. Epub 2018 May 3.
9
Polyketide mimetics yield structural and mechanistic insights into product template domain function in nonreducing polyketide synthases.
Proc Natl Acad Sci U S A. 2017 May 23;114(21):E4142-E4148. doi: 10.1073/pnas.1609001114. Epub 2017 May 8.
10
Resin supported acyl carrier protein labeling strategies.
RSC Adv. 2014;4(18):9092-9097. doi: 10.1039/C3RA47847E.
本文引用的文献
1
Mechanism-based protein cross-linking probes to investigate carrier protein-mediated biosynthesis.
ACS Chem Biol. 2006 Dec 20;1(11):687-91. doi: 10.1021/cb6003965.
2
Synthesis and evaluation of bioorthogonal pantetheine analogues for in vivo protein modification.
J Am Chem Soc. 2006 Sep 20;128(37):12174-84. doi: 10.1021/ja063217n.
3
Complete set of ORF clones of Escherichia coli ASKA library (a complete set of E. coli K-12 ORF archive): unique resources for biological research.
DNA Res. 2005;12(5):291-9. doi: 10.1093/dnares/dsi012. Epub 2006 Jan 9.
4
Architecture of a fungal fatty acid synthase at 5 A resolution.
Science. 2006 Mar 3;311(5765):1263-7. doi: 10.1126/science.1123251.
5
Architecture of mammalian fatty acid synthase at 4.5 A resolution.
Science. 2006 Mar 3;311(5765):1258-62. doi: 10.1126/science.1123248.
6
One-pot chemo-enzymatic synthesis of reporter-modified proteins.
Org Biomol Chem. 2006 Jan 7;4(1):44-6. doi: 10.1039/b512735a. Epub 2005 Nov 24.
7
In vivo reporter labeling of proteins via metabolic delivery of coenzyme A analogues.
J Am Chem Soc. 2005 Aug 17;127(32):11234-5. doi: 10.1021/ja052911k.
8
Proteome-scale purification of human proteins from bacteria.
Proc Natl Acad Sci U S A. 2002 Mar 5;99(5):2654-9. doi: 10.1073/pnas.042684199.
9
Identification and analysis of the acyl carrier protein (ACP) docking site on beta-ketoacyl-ACP synthase III.
J Biol Chem. 2001 Mar 16;276(11):8231-8. doi: 10.1074/jbc.M008042200. Epub 2000 Nov 14.
10
Crystal structure of beta-ketoacyl-acyl carrier protein synthase III. A key condensing enzyme in bacterial fatty acid biosynthesis.
J Biol Chem. 1999 Dec 17;274(51):36465-71. doi: 10.1074/jbc.274.51.36465.
Zotero 插件