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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ftcdlib:oai:escholarship.org/ark:/13030/qt3mm7q8m2 2023-05-15T17:58:17+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 e17624 2011-03-18 application/pdf https://escholarship.org/uc/item/3mm7q8m2 unknown eScholarship, University of California qt3mm7q8m2 https://escholarship.org/uc/item/3mm7q8m2 public PloS one, vol 6, iss 3 Cell Wall Psychrobacter Peptide Synthases Crystallography X-Ray Amino Acid Sequence Protein Conformation Sequence Homology Amino Acid Structure-Activity Relationship Substrate Specificity Models Molecular Molecular Sequence Data General Science & Technology article 2011 ftcdlib 2020-06-06T07:53:16Z 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 University of California: eScholarship |
institution |
Open Polar |
collection |
University of California: eScholarship |
op_collection_id |
ftcdlib |
language |
unknown |
topic |
Cell Wall Psychrobacter Peptide Synthases Crystallography X-Ray Amino Acid Sequence Protein Conformation Sequence Homology Amino Acid Structure-Activity Relationship Substrate Specificity Models Molecular Molecular Sequence Data General Science & Technology |
spellingShingle |
Cell Wall Psychrobacter Peptide Synthases Crystallography X-Ray Amino Acid Sequence Protein Conformation Sequence Homology Amino Acid Structure-Activity Relationship Substrate Specificity Models Molecular Molecular Sequence Data General Science & Technology 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. |
topic_facet |
Cell Wall Psychrobacter Peptide Synthases Crystallography X-Ray Amino Acid Sequence Protein Conformation Sequence Homology Amino Acid Structure-Activity Relationship Substrate Specificity Models Molecular Molecular Sequence Data General Science & Technology |
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 |
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 |
eScholarship, University of California |
publishDate |
2011 |
url |
https://escholarship.org/uc/item/3mm7q8m2 |
op_coverage |
e17624 |
genre |
permafrost |
genre_facet |
permafrost |
op_source |
PloS one, vol 6, iss 3 |
op_relation |
qt3mm7q8m2 https://escholarship.org/uc/item/3mm7q8m2 |
op_rights |
public |
_version_ |
1766166857135423488 |