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Conversion of an amide to a high-energy thioester by Staphylococcus aureus sortase A is powered by variable binding affinity for calcium

Wang, Xiao; Chen, Jia-Liang; Otting, Gottfried; Su, Xun-Cheng

Description

Thioesters are key intermediates in biology, which often are generated from less energy-rich amide precursors. Staphylococcus aureus sortase A (SrtA) is an enzyme widely used in biotechnology for peptide ligation. The reaction proceeds in two steps, where the first step involves the conversion of an amide bond of substrate peptide into a thioester intermediate with the enzyme. Here we show that the free energy required for this step is matched by an about 30-fold increase in binding affinity of...[Show more]

dc.contributor.advisorhttp://creativecommons.org/licenses/by/4.0/
dc.contributor.authorWang, Xiao
dc.contributor.authorChen, Jia-Liang
dc.contributor.authorOtting, Gottfried
dc.contributor.authorSu, Xun-Cheng
dc.date.accessioned2019-04-13T02:16:58Z
dc.date.available2019-04-13T02:16:58Z
dc.identifier.issn2045-2322
dc.identifier.urihttp://hdl.handle.net/1885/159554
dc.description.abstractThioesters are key intermediates in biology, which often are generated from less energy-rich amide precursors. Staphylococcus aureus sortase A (SrtA) is an enzyme widely used in biotechnology for peptide ligation. The reaction proceeds in two steps, where the first step involves the conversion of an amide bond of substrate peptide into a thioester intermediate with the enzyme. Here we show that the free energy required for this step is matched by an about 30-fold increase in binding affinity of a calcium ion at the calcium binding site of SrtA, which is remote from the thioester bond. The magnitude of this allosteric effect highlights the importance of calcium for the activity of SrtA. The increase in calcium binding affinity upon binding of substrate not only achieves catalytic formation of an energy-rich intermediate in the absence of nucleotide triphosphates or any tight non-covalent enzyme-substrate interactions, but is also accompanied by accumulation of the labile thioester intermediate, which makes it directly observable in nuclear magnetic resonance (NMR) spectra.
dc.format.mimetypeapplication/pdf
dc.language.isoen_AU
dc.publisherNature Publishing Group
dc.rightsAuthor/s retain copyright
dc.sourceScientific Reports
dc.titleConversion of an amide to a high-energy thioester by Staphylococcus aureus sortase A is powered by variable binding affinity for calcium
dc.typeJournal article
local.description.notesImported from ARIES
local.identifier.citationvolume8
dc.date.issued2018
local.identifier.absfor030606 - Structural Chemistry and Spectroscopy
local.identifier.absfor030406 - Proteins and Peptides
local.identifier.absfor030403 - Characterisation of Biological Macromolecules
local.identifier.ariespublicationu3102795xPUB66
local.type.statusPublished Version
local.contributor.affiliationWang, Xiao, Nankai University
local.contributor.affiliationChen, Jia-Liang, Nankai University
local.contributor.affiliationOtting, Gottfried, College of Science, ANU
local.contributor.affiliationSu, Xun-Cheng, Nankai University
local.bibliographicCitation.issue16371
local.identifier.doi10.1038/s41598-018-34752-6
local.identifier.absseo970106 - Expanding Knowledge in the Biological Sciences
dc.date.updated2019-03-12T07:25:18Z
local.identifier.scopusID2-s2.0-85056131350
dcterms.accessRightsOpen Access
dc.provenanceJournal: Scientific Reports (ESSN: 2045-2322) RoMEO: This is a RoMEO green journal Listed in: DOAJ as an open access journal Author's Pre-print: green tick author can archive pre-print (ie pre-refereeing) Author's Post-print: green tick author can archive post-print (ie final draft post-refereeing) Publisher's Version/PDF: green tick author can archive publisher's version/PDF General Conditions: Author's pre-print on any website (Research articles only) Immediately upon publication On any website Publisher's version/PDF may be used Must link to publisher version Publisher copyright and source must be acknowledged and DOI cited Authors retain copyright
dc.rights.licenseThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Cre- ative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not per- mitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
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