Enzymatic synthesis of c-di-GMP using a thermophilic diguanylate cyclase - PubMed (original) (raw)
. 2009 Jun 15;389(2):138-42.
doi: 10.1016/j.ab.2009.03.031. Epub 2009 Mar 27.
Affiliations
- PMID: 19328769
- DOI: 10.1016/j.ab.2009.03.031
Enzymatic synthesis of c-di-GMP using a thermophilic diguanylate cyclase
Feng Rao et al. Anal Biochem. 2009.
Abstract
The cyclic dinucleotide c-di-GMP is a widespread bacterial messenger molecule with potential application as a therapeutic agent for treating bacterial infection. Current enzymatic synthesis of c-di-GMP using mesophilic diguanylate cyclase (DGC) proteins suffers from low production yield due to protein instability and strong product inhibition. Here we report the overexpression and characterization of a stand-alone thermophilic diguanylate cyclase domain (tDGC) protein with enhanced thermostability. The product inhibition that severely limited production yield was significantly alleviated by mutation of a conserved residue in the putative regulatory I-site. With the mutant tDGC, we demonstrated that hundreds of milligrams of c-di-GMP can be readily prepared by using the optimized procedures for enzymatic reaction and product purification. The thermophilic enzyme will be a valuable tool for other research laboratories for c-di-GMP synthesis as well as the preparation of c-di-GMP derivatives.
Similar articles
- Enzymatic Production of c-di-GMP Using a Thermophilic Diguanylate Cyclase.
Venkataramani P, Liang ZX. Venkataramani P, et al. Methods Mol Biol. 2017;1657:11-22. doi: 10.1007/978-1-4939-7240-1_2. Methods Mol Biol. 2017. PMID: 28889282 - Structure of a diguanylate cyclase from Thermotoga maritima: insights into activation, feedback inhibition and thermostability.
Deepthi A, Liew CW, Liang ZX, Swaminathan K, Lescar J. Deepthi A, et al. PLoS One. 2014 Oct 31;9(10):e110912. doi: 10.1371/journal.pone.0110912. eCollection 2014. PLoS One. 2014. PMID: 25360685 Free PMC article. - Enzymatic synthesis of 2'-ara and 2'-deoxy analogues of c-di-GMP.
Shchokolova AS, Rymko AN, Kvach SV, Shabunya PS, Fatykhava SA, Zinchenko AI. Shchokolova AS, et al. Nucleosides Nucleotides Nucleic Acids. 2015;34(6):416-23. doi: 10.1080/15257770.2015.1006775. Nucleosides Nucleotides Nucleic Acids. 2015. PMID: 25965330 - [Activity of cyclic diguanylate (c-di-GMP) in bacteria and the study of its derivatives].
Na LX, Yang ZJ. Na LX, et al. Yao Xue Xue Bao. 2012 Mar;47(3):307-12. Yao Xue Xue Bao. 2012. PMID: 22645753 Review. Chinese. - 3',5'-Cyclic diguanylic acid: a small nucleotide that makes big impacts.
Yan H, Chen W. Yan H, et al. Chem Soc Rev. 2010 Aug;39(8):2914-24. doi: 10.1039/b914942m. Epub 2010 Jun 25. Chem Soc Rev. 2010. PMID: 20577685 Review.
Cited by
- cGAS produces a 2'-5'-linked cyclic dinucleotide second messenger that activates STING.
Ablasser A, Goldeck M, Cavlar T, Deimling T, Witte G, Röhl I, Hopfner KP, Ludwig J, Hornung V. Ablasser A, et al. Nature. 2013 Jun 20;498(7454):380-4. doi: 10.1038/nature12306. Epub 2013 May 30. Nature. 2013. PMID: 23722158 Free PMC article. - Progress in Understanding the Molecular Basis Underlying Functional Diversification of Cyclic Dinucleotide Turnover Proteins.
Römling U, Liang ZX, Dow JM. Römling U, et al. J Bacteriol. 2017 Feb 14;199(5):e00790-16. doi: 10.1128/JB.00790-16. Print 2017 Mar 1. J Bacteriol. 2017. PMID: 28031279 Free PMC article. Review. - The World of Cyclic Dinucleotides in Bacterial Behavior.
Aline Dias da P, Nathalia Marins de A, Gabriel Guarany de A, Robson Francisco de S, Cristiane Rodrigues G. Aline Dias da P, et al. Molecules. 2020 May 25;25(10):2462. doi: 10.3390/molecules25102462. Molecules. 2020. PMID: 32466317 Free PMC article. Review. - Crystallization and preliminary X-ray diffraction studies of Xanthomonas campestris PNPase in the presence of c-di-GMP.
Wang YC, Chin KH, Chuah ML, Liang ZX, Chou SH. Wang YC, et al. Acta Crystallogr Sect F Struct Biol Cryst Commun. 2012 Oct 1;68(Pt 10):1247-50. doi: 10.1107/S1744309112036202. Epub 2012 Sep 29. Acta Crystallogr Sect F Struct Biol Cryst Commun. 2012. PMID: 23027759 Free PMC article. - Identification of small molecules inhibiting diguanylate cyclases to control bacterial biofilm development.
Sambanthamoorthy K, Luo C, Pattabiraman N, Feng X, Koestler B, Waters CM, Palys TJ. Sambanthamoorthy K, et al. Biofouling. 2014 Jan;30(1):17-28. doi: 10.1080/08927014.2013.832224. Epub 2013 Oct 11. Biofouling. 2014. PMID: 24117391 Free PMC article.
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources