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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ftpubmed:oai:pubmedcentral.nih.gov:3060825 2023-05-15T17:58:09+02:00 Structure and Function of the First Full-Length Murein Peptide Ligase (Mpl) Cell Wall Recycling Protein Das, Debanu Hervé, Mireille Feuerhelm, Julie Farr, Carol L. Chiu, Hsiu-Ju Elsliger, Marc-André Knuth, Mark W. Klock, Heath E. Miller, Mitchell D. Godzik, Adam Lesley, Scott A. Deacon, Ashley M. Mengin-Lecreulx, Dominique Wilson, Ian A. 2011-03-18 http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3060825 http://www.ncbi.nlm.nih.gov/pubmed/21445265 https://doi.org/10.1371/journal.pone.0017624 en eng Public Library of Science http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3060825 http://www.ncbi.nlm.nih.gov/pubmed/21445265 http://dx.doi.org/10.1371/journal.pone.0017624 Das et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. CC-BY Research Article Text 2011 ftpubmed https://doi.org/10.1371/journal.pone.0017624 2013-09-03T12:26:28Z 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. Text permafrost PubMed Central (PMC) PLoS ONE 6 3 e17624 |
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Research Article |
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Research Article Das, Debanu Hervé, Mireille Feuerhelm, Julie Farr, Carol L. Chiu, Hsiu-Ju Elsliger, Marc-André Knuth, Mark W. Klock, Heath E. Miller, Mitchell D. Godzik, Adam Lesley, Scott A. Deacon, Ashley M. Mengin-Lecreulx, Dominique Wilson, Ian A. Structure and Function of the First Full-Length Murein Peptide Ligase (Mpl) Cell Wall Recycling Protein |
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Research Article |
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 |
Text |
author |
Das, Debanu Hervé, Mireille Feuerhelm, Julie Farr, Carol L. Chiu, Hsiu-Ju Elsliger, Marc-André Knuth, Mark W. Klock, Heath E. Miller, Mitchell D. Godzik, Adam Lesley, Scott A. Deacon, Ashley M. Mengin-Lecreulx, Dominique Wilson, Ian A. |
author_facet |
Das, Debanu Hervé, Mireille Feuerhelm, Julie Farr, Carol L. Chiu, Hsiu-Ju Elsliger, Marc-André Knuth, Mark W. Klock, Heath E. Miller, Mitchell D. Godzik, Adam Lesley, Scott A. Deacon, Ashley M. Mengin-Lecreulx, Dominique Wilson, Ian A. |
author_sort |
Das, Debanu |
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 |
publishDate |
2011 |
url |
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3060825 http://www.ncbi.nlm.nih.gov/pubmed/21445265 https://doi.org/10.1371/journal.pone.0017624 |
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permafrost |
genre_facet |
permafrost |
op_relation |
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3060825 http://www.ncbi.nlm.nih.gov/pubmed/21445265 http://dx.doi.org/10.1371/journal.pone.0017624 |
op_rights |
Das et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
op_rightsnorm |
CC-BY |
op_doi |
https://doi.org/10.1371/journal.pone.0017624 |
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PLoS ONE |
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6 |
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3 |
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e17624 |
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1766166690459025408 |