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Novel Applications of Biocatalysis to Stereochemistry Determination of 2′3′-cGAMP Bisphosphorothioate (2′3′-cG(S)A(S)MP)

[Image: see text] The metazoan second messenger 2′3′-cyclic guanosine monophosphate–adenosine monophosphate (cGAMP) is a cyclic dinucleotide (CDN) that induces secretion of type I interferons and activates the immune system and has thus attracted significant interest as a vaccine adjuvant or immunot...

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Autores principales: Lim, Jongwon, Kim, Hai-Young
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Chemical Society 2020
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7301576/
https://www.ncbi.nlm.nih.gov/pubmed/32566885
http://dx.doi.org/10.1021/acsomega.0c01942
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author Lim, Jongwon
Kim, Hai-Young
author_facet Lim, Jongwon
Kim, Hai-Young
author_sort Lim, Jongwon
collection PubMed
description [Image: see text] The metazoan second messenger 2′3′-cyclic guanosine monophosphate–adenosine monophosphate (cGAMP) is a cyclic dinucleotide (CDN) that induces secretion of type I interferons and activates the immune system and has thus attracted significant interest as a vaccine adjuvant or immunotherapeutic. CDN bisphosphorothioates are of particular interest because of their increased hydrolytic stability and improved cell activities. In our work with CDN bisphosphorothioates, we sought a method for systematic determination of the absolute stereochemistry of their phosphorothioate stereocenters. A novel biocatalytic method employing snake venom phosphodiesterase (svPDE) and nP1 has been developed and successfully applied to stereochemistry determination of 2′3′-cGAMP bisphosphorothioates. This method unambiguously assigned the phosphorothioate stereochemistry of four diastereomers of 2′3′-cGAMP bisphosphorothioate by analyzing distinct hydrolysis patterns of the bisphosphorothioate diastereomers upon incubation with svPDE and nP1. Furthermore, the regiospecificity as well as stereospecificity of both svPDE and nP1 toward 2′3′-cGAMP bisphosphorothioate has been elucidated.
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spelling pubmed-73015762020-06-19 Novel Applications of Biocatalysis to Stereochemistry Determination of 2′3′-cGAMP Bisphosphorothioate (2′3′-cG(S)A(S)MP) Lim, Jongwon Kim, Hai-Young ACS Omega [Image: see text] The metazoan second messenger 2′3′-cyclic guanosine monophosphate–adenosine monophosphate (cGAMP) is a cyclic dinucleotide (CDN) that induces secretion of type I interferons and activates the immune system and has thus attracted significant interest as a vaccine adjuvant or immunotherapeutic. CDN bisphosphorothioates are of particular interest because of their increased hydrolytic stability and improved cell activities. In our work with CDN bisphosphorothioates, we sought a method for systematic determination of the absolute stereochemistry of their phosphorothioate stereocenters. A novel biocatalytic method employing snake venom phosphodiesterase (svPDE) and nP1 has been developed and successfully applied to stereochemistry determination of 2′3′-cGAMP bisphosphorothioates. This method unambiguously assigned the phosphorothioate stereochemistry of four diastereomers of 2′3′-cGAMP bisphosphorothioate by analyzing distinct hydrolysis patterns of the bisphosphorothioate diastereomers upon incubation with svPDE and nP1. Furthermore, the regiospecificity as well as stereospecificity of both svPDE and nP1 toward 2′3′-cGAMP bisphosphorothioate has been elucidated. American Chemical Society 2020-06-03 /pmc/articles/PMC7301576/ /pubmed/32566885 http://dx.doi.org/10.1021/acsomega.0c01942 Text en Copyright © 2020 American Chemical Society This is an open access article published under an ACS AuthorChoice License (http://pubs.acs.org/page/policy/authorchoice_termsofuse.html) , which permits copying and redistribution of the article or any adaptations for non-commercial purposes.
spellingShingle Lim, Jongwon
Kim, Hai-Young
Novel Applications of Biocatalysis to Stereochemistry Determination of 2′3′-cGAMP Bisphosphorothioate (2′3′-cG(S)A(S)MP)
title Novel Applications of Biocatalysis to Stereochemistry Determination of 2′3′-cGAMP Bisphosphorothioate (2′3′-cG(S)A(S)MP)
title_full Novel Applications of Biocatalysis to Stereochemistry Determination of 2′3′-cGAMP Bisphosphorothioate (2′3′-cG(S)A(S)MP)
title_fullStr Novel Applications of Biocatalysis to Stereochemistry Determination of 2′3′-cGAMP Bisphosphorothioate (2′3′-cG(S)A(S)MP)
title_full_unstemmed Novel Applications of Biocatalysis to Stereochemistry Determination of 2′3′-cGAMP Bisphosphorothioate (2′3′-cG(S)A(S)MP)
title_short Novel Applications of Biocatalysis to Stereochemistry Determination of 2′3′-cGAMP Bisphosphorothioate (2′3′-cG(S)A(S)MP)
title_sort novel applications of biocatalysis to stereochemistry determination of 2′3′-cgamp bisphosphorothioate (2′3′-cg(s)a(s)mp)
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7301576/
https://www.ncbi.nlm.nih.gov/pubmed/32566885
http://dx.doi.org/10.1021/acsomega.0c01942
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