Structure and function of the first full-length murein peptide ligase (Mpl) cell wall recycling protein.
Bacterial cell walls contain peptidoglycan, an essential polymer made by enzymes in the Mur pathway. These proteins are specific to bacteria, which make them targets for drug discovery. MurC, MurD, MurE and MurF catalyze the synthesis of the peptidoglycan precursor UDP-N-acetylmuramoyl-L-alanyl-γ-D-...
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ftdoajarticles:oai:doaj.org/article:3579f29d15a044a7b200ed31f29f9110 2023-05-15T17:58:09+02:00 Structure and function of the first full-length murein peptide ligase (Mpl) cell wall recycling protein. Debanu Das Mireille Hervé Julie Feuerhelm Carol L Farr Hsiu-Ju Chiu Marc-André Elsliger Mark W Knuth Heath E Klock Mitchell D Miller Adam Godzik Scott A Lesley Ashley M Deacon Dominique Mengin-Lecreulx Ian A Wilson 2011-03-01T00:00:00Z https://doi.org/10.1371/journal.pone.0017624 https://doaj.org/article/3579f29d15a044a7b200ed31f29f9110 EN eng Public Library of Science (PLoS) http://europepmc.org/articles/PMC3060825?pdf=render https://doaj.org/toc/1932-6203 1932-6203 doi:10.1371/journal.pone.0017624 https://doaj.org/article/3579f29d15a044a7b200ed31f29f9110 PLoS ONE, Vol 6, Iss 3, p e17624 (2011) Medicine R Science Q article 2011 ftdoajarticles https://doi.org/10.1371/journal.pone.0017624 2022-12-31T16:30:20Z Bacterial cell walls contain peptidoglycan, an essential polymer made by enzymes in the Mur pathway. These proteins are specific to bacteria, which make them targets for drug discovery. MurC, MurD, MurE and MurF catalyze the synthesis of the peptidoglycan precursor UDP-N-acetylmuramoyl-L-alanyl-γ-D-glutamyl-meso-diaminopimelyl-D-alanyl-D-alanine by the sequential addition of amino acids onto UDP-N-acetylmuramic acid (UDP-MurNAc). MurC-F enzymes have been extensively studied by biochemistry and X-ray crystallography. In gram-negative bacteria, ∼30-60% of the bacterial cell wall is recycled during each generation. Part of this recycling process involves the murein peptide ligase (Mpl), which attaches the breakdown product, the tripeptide L-alanyl-γ-D-glutamyl-meso-diaminopimelate, to UDP-MurNAc. We present the crystal structure at 1.65 Å resolution of a full-length Mpl from the permafrost bacterium Psychrobacter arcticus 273-4 (PaMpl). Although the Mpl structure has similarities to Mur enzymes, it has unique sequence and structure features that are likely related to its role in cell wall recycling, a function that differentiates it from the MurC-F enzymes. We have analyzed the sequence-structure relationships that are unique to Mpl proteins and compared them to MurC-F ligases. We have also characterized the biochemical properties of this enzyme (optimal temperature, pH and magnesium binding profiles and kinetic parameters). Although the structure does not contain any bound substrates, we have identified ∼30 residues that are likely to be important for recognition of the tripeptide and UDP-MurNAc substrates, as well as features that are unique to Psychrobacter Mpl proteins. These results provide the basis for future mutational studies for more extensive function characterization of the Mpl sequence-structure relationships. Article in Journal/Newspaper permafrost Directory of Open Access Journals: DOAJ Articles PLoS ONE 6 3 e17624 |
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Directory of Open Access Journals: DOAJ Articles |
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ftdoajarticles |
language |
English |
topic |
Medicine R Science Q |
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Medicine R Science Q Debanu Das Mireille Hervé Julie Feuerhelm Carol L Farr Hsiu-Ju Chiu Marc-André Elsliger Mark W Knuth Heath E Klock Mitchell D Miller Adam Godzik Scott A Lesley Ashley M Deacon Dominique Mengin-Lecreulx Ian A Wilson Structure and function of the first full-length murein peptide ligase (Mpl) cell wall recycling protein. |
topic_facet |
Medicine R Science Q |
description |
Bacterial cell walls contain peptidoglycan, an essential polymer made by enzymes in the Mur pathway. These proteins are specific to bacteria, which make them targets for drug discovery. MurC, MurD, MurE and MurF catalyze the synthesis of the peptidoglycan precursor UDP-N-acetylmuramoyl-L-alanyl-γ-D-glutamyl-meso-diaminopimelyl-D-alanyl-D-alanine by the sequential addition of amino acids onto UDP-N-acetylmuramic acid (UDP-MurNAc). MurC-F enzymes have been extensively studied by biochemistry and X-ray crystallography. In gram-negative bacteria, ∼30-60% of the bacterial cell wall is recycled during each generation. Part of this recycling process involves the murein peptide ligase (Mpl), which attaches the breakdown product, the tripeptide L-alanyl-γ-D-glutamyl-meso-diaminopimelate, to UDP-MurNAc. We present the crystal structure at 1.65 Å resolution of a full-length Mpl from the permafrost bacterium Psychrobacter arcticus 273-4 (PaMpl). Although the Mpl structure has similarities to Mur enzymes, it has unique sequence and structure features that are likely related to its role in cell wall recycling, a function that differentiates it from the MurC-F enzymes. We have analyzed the sequence-structure relationships that are unique to Mpl proteins and compared them to MurC-F ligases. We have also characterized the biochemical properties of this enzyme (optimal temperature, pH and magnesium binding profiles and kinetic parameters). Although the structure does not contain any bound substrates, we have identified ∼30 residues that are likely to be important for recognition of the tripeptide and UDP-MurNAc substrates, as well as features that are unique to Psychrobacter Mpl proteins. These results provide the basis for future mutational studies for more extensive function characterization of the Mpl sequence-structure relationships. |
format |
Article in Journal/Newspaper |
author |
Debanu Das Mireille Hervé Julie Feuerhelm Carol L Farr Hsiu-Ju Chiu Marc-André Elsliger Mark W Knuth Heath E Klock Mitchell D Miller Adam Godzik Scott A Lesley Ashley M Deacon Dominique Mengin-Lecreulx Ian A Wilson |
author_facet |
Debanu Das Mireille Hervé Julie Feuerhelm Carol L Farr Hsiu-Ju Chiu Marc-André Elsliger Mark W Knuth Heath E Klock Mitchell D Miller Adam Godzik Scott A Lesley Ashley M Deacon Dominique Mengin-Lecreulx Ian A Wilson |
author_sort |
Debanu Das |
title |
Structure and function of the first full-length murein peptide ligase (Mpl) cell wall recycling protein. |
title_short |
Structure and function of the first full-length murein peptide ligase (Mpl) cell wall recycling protein. |
title_full |
Structure and function of the first full-length murein peptide ligase (Mpl) cell wall recycling protein. |
title_fullStr |
Structure and function of the first full-length murein peptide ligase (Mpl) cell wall recycling protein. |
title_full_unstemmed |
Structure and function of the first full-length murein peptide ligase (Mpl) cell wall recycling protein. |
title_sort |
structure and function of the first full-length murein peptide ligase (mpl) cell wall recycling protein. |
publisher |
Public Library of Science (PLoS) |
publishDate |
2011 |
url |
https://doi.org/10.1371/journal.pone.0017624 https://doaj.org/article/3579f29d15a044a7b200ed31f29f9110 |
genre |
permafrost |
genre_facet |
permafrost |
op_source |
PLoS ONE, Vol 6, Iss 3, p e17624 (2011) |
op_relation |
http://europepmc.org/articles/PMC3060825?pdf=render https://doaj.org/toc/1932-6203 1932-6203 doi:10.1371/journal.pone.0017624 https://doaj.org/article/3579f29d15a044a7b200ed31f29f9110 |
op_doi |
https://doi.org/10.1371/journal.pone.0017624 |
container_title |
PLoS ONE |
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6 |
container_issue |
3 |
container_start_page |
e17624 |
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1766166699393941504 |