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A cryptic third active site in cyanophycin synthetase creates primers for polymerization

Cyanophycin is a nitrogen reserve biopolymer in many bacteria that has promising industrial applications. Made by cyanophycin synthetase 1 (CphA1), it has a poly-L-Asp backbone with L-Arg residues attached to each aspartate sidechain. CphA1s are thought to typically require existing segments of cyan...

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Autores principales: Sharon, Itai, Pinus, Sharon, Grogg, Marcel, Moitessier, Nicolas, Hilvert, Donald, Schmeing, T. Martin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9262961/
https://www.ncbi.nlm.nih.gov/pubmed/35798723
http://dx.doi.org/10.1038/s41467-022-31542-7
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author Sharon, Itai
Pinus, Sharon
Grogg, Marcel
Moitessier, Nicolas
Hilvert, Donald
Schmeing, T. Martin
author_facet Sharon, Itai
Pinus, Sharon
Grogg, Marcel
Moitessier, Nicolas
Hilvert, Donald
Schmeing, T. Martin
author_sort Sharon, Itai
collection PubMed
description Cyanophycin is a nitrogen reserve biopolymer in many bacteria that has promising industrial applications. Made by cyanophycin synthetase 1 (CphA1), it has a poly-L-Asp backbone with L-Arg residues attached to each aspartate sidechain. CphA1s are thought to typically require existing segments of cyanophycin to act as primers for cyanophycin polymerization. In this study, we show that most CphA1s will not require exogenous primers and discover the surprising cause of primer independence: CphA1 can make minute quantities of cyanophycin without primer, and an unexpected, cryptic metallopeptidase-like active site in the N-terminal domain of many CphA1s digests these into primers, solving the problem of primer availability. We present co-complex cryo-EM structures, make mutations that transition CphA1s between primer dependence and independence, and demonstrate that primer dependence can be a limiting factor for cyanophycin production in heterologous hosts. In CphA1, domains with opposite catalytic activities combine into a remarkable, self-sufficient, biosynthetic nanomachine.
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spelling pubmed-92629612022-07-09 A cryptic third active site in cyanophycin synthetase creates primers for polymerization Sharon, Itai Pinus, Sharon Grogg, Marcel Moitessier, Nicolas Hilvert, Donald Schmeing, T. Martin Nat Commun Article Cyanophycin is a nitrogen reserve biopolymer in many bacteria that has promising industrial applications. Made by cyanophycin synthetase 1 (CphA1), it has a poly-L-Asp backbone with L-Arg residues attached to each aspartate sidechain. CphA1s are thought to typically require existing segments of cyanophycin to act as primers for cyanophycin polymerization. In this study, we show that most CphA1s will not require exogenous primers and discover the surprising cause of primer independence: CphA1 can make minute quantities of cyanophycin without primer, and an unexpected, cryptic metallopeptidase-like active site in the N-terminal domain of many CphA1s digests these into primers, solving the problem of primer availability. We present co-complex cryo-EM structures, make mutations that transition CphA1s between primer dependence and independence, and demonstrate that primer dependence can be a limiting factor for cyanophycin production in heterologous hosts. In CphA1, domains with opposite catalytic activities combine into a remarkable, self-sufficient, biosynthetic nanomachine. Nature Publishing Group UK 2022-07-07 /pmc/articles/PMC9262961/ /pubmed/35798723 http://dx.doi.org/10.1038/s41467-022-31542-7 Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open Access This 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 Creative 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 permitted 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/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Sharon, Itai
Pinus, Sharon
Grogg, Marcel
Moitessier, Nicolas
Hilvert, Donald
Schmeing, T. Martin
A cryptic third active site in cyanophycin synthetase creates primers for polymerization
title A cryptic third active site in cyanophycin synthetase creates primers for polymerization
title_full A cryptic third active site in cyanophycin synthetase creates primers for polymerization
title_fullStr A cryptic third active site in cyanophycin synthetase creates primers for polymerization
title_full_unstemmed A cryptic third active site in cyanophycin synthetase creates primers for polymerization
title_short A cryptic third active site in cyanophycin synthetase creates primers for polymerization
title_sort cryptic third active site in cyanophycin synthetase creates primers for polymerization
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9262961/
https://www.ncbi.nlm.nih.gov/pubmed/35798723
http://dx.doi.org/10.1038/s41467-022-31542-7
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