Cargando…

Identification and structural characterization of LytU, a unique peptidoglycan endopeptidase from the lysostaphin family

We introduce LytU, a short member of the lysostaphin family of zinc-dependent pentaglycine endopeptidases. It is a potential antimicrobial agent for S. aureus infections and its gene transcription is highly upregulated upon antibiotic treatments along with other genes involved in cell wall synthesis...

Descripción completa

Detalles Bibliográficos
Autores principales: Raulinaitis, Vytas, Tossavainen, Helena, Aitio, Olli, Juuti, Jarmo T., Hiramatsu, Keiichi, Kontinen, Vesa, Permi, Perttu
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5519744/
https://www.ncbi.nlm.nih.gov/pubmed/28729697
http://dx.doi.org/10.1038/s41598-017-06135-w
_version_ 1783251684788535296
author Raulinaitis, Vytas
Tossavainen, Helena
Aitio, Olli
Juuti, Jarmo T.
Hiramatsu, Keiichi
Kontinen, Vesa
Permi, Perttu
author_facet Raulinaitis, Vytas
Tossavainen, Helena
Aitio, Olli
Juuti, Jarmo T.
Hiramatsu, Keiichi
Kontinen, Vesa
Permi, Perttu
author_sort Raulinaitis, Vytas
collection PubMed
description We introduce LytU, a short member of the lysostaphin family of zinc-dependent pentaglycine endopeptidases. It is a potential antimicrobial agent for S. aureus infections and its gene transcription is highly upregulated upon antibiotic treatments along with other genes involved in cell wall synthesis. We found this enzyme to be responsible for the opening of the cell wall peptidoglycan layer during cell divisions in S. aureus. LytU is anchored in the plasma membrane with the active part residing in the periplasmic space. It has a unique Ile/Lys insertion at position 151 that resides in the catalytic site-neighbouring loop and is vital for the enzymatic activity but not affecting the overall structure common to the lysostaphin family. Purified LytU lyses S. aureus cells and cleaves pentaglycine, a reaction conveniently monitored by NMR spectroscopy. Substituting the cofactor zinc ion with a copper or cobalt ion remarkably increases the rate of pentaglycine cleavage. NMR and isothermal titration calorimetry further reveal that, uniquely for its family, LytU is able to bind a second zinc ion which is coordinated by catalytic histidines and is therefore inhibitory. The pH-dependence and high affinity of binding carry further physiological implications.
format Online
Article
Text
id pubmed-5519744
institution National Center for Biotechnology Information
language English
publishDate 2017
publisher Nature Publishing Group UK
record_format MEDLINE/PubMed
spelling pubmed-55197442017-07-26 Identification and structural characterization of LytU, a unique peptidoglycan endopeptidase from the lysostaphin family Raulinaitis, Vytas Tossavainen, Helena Aitio, Olli Juuti, Jarmo T. Hiramatsu, Keiichi Kontinen, Vesa Permi, Perttu Sci Rep Article We introduce LytU, a short member of the lysostaphin family of zinc-dependent pentaglycine endopeptidases. It is a potential antimicrobial agent for S. aureus infections and its gene transcription is highly upregulated upon antibiotic treatments along with other genes involved in cell wall synthesis. We found this enzyme to be responsible for the opening of the cell wall peptidoglycan layer during cell divisions in S. aureus. LytU is anchored in the plasma membrane with the active part residing in the periplasmic space. It has a unique Ile/Lys insertion at position 151 that resides in the catalytic site-neighbouring loop and is vital for the enzymatic activity but not affecting the overall structure common to the lysostaphin family. Purified LytU lyses S. aureus cells and cleaves pentaglycine, a reaction conveniently monitored by NMR spectroscopy. Substituting the cofactor zinc ion with a copper or cobalt ion remarkably increases the rate of pentaglycine cleavage. NMR and isothermal titration calorimetry further reveal that, uniquely for its family, LytU is able to bind a second zinc ion which is coordinated by catalytic histidines and is therefore inhibitory. The pH-dependence and high affinity of binding carry further physiological implications. Nature Publishing Group UK 2017-07-20 /pmc/articles/PMC5519744/ /pubmed/28729697 http://dx.doi.org/10.1038/s41598-017-06135-w Text en © The Author(s) 2017 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/.
spellingShingle Article
Raulinaitis, Vytas
Tossavainen, Helena
Aitio, Olli
Juuti, Jarmo T.
Hiramatsu, Keiichi
Kontinen, Vesa
Permi, Perttu
Identification and structural characterization of LytU, a unique peptidoglycan endopeptidase from the lysostaphin family
title Identification and structural characterization of LytU, a unique peptidoglycan endopeptidase from the lysostaphin family
title_full Identification and structural characterization of LytU, a unique peptidoglycan endopeptidase from the lysostaphin family
title_fullStr Identification and structural characterization of LytU, a unique peptidoglycan endopeptidase from the lysostaphin family
title_full_unstemmed Identification and structural characterization of LytU, a unique peptidoglycan endopeptidase from the lysostaphin family
title_short Identification and structural characterization of LytU, a unique peptidoglycan endopeptidase from the lysostaphin family
title_sort identification and structural characterization of lytu, a unique peptidoglycan endopeptidase from the lysostaphin family
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5519744/
https://www.ncbi.nlm.nih.gov/pubmed/28729697
http://dx.doi.org/10.1038/s41598-017-06135-w
work_keys_str_mv AT raulinaitisvytas identificationandstructuralcharacterizationoflytuauniquepeptidoglycanendopeptidasefromthelysostaphinfamily
AT tossavainenhelena identificationandstructuralcharacterizationoflytuauniquepeptidoglycanendopeptidasefromthelysostaphinfamily
AT aitioolli identificationandstructuralcharacterizationoflytuauniquepeptidoglycanendopeptidasefromthelysostaphinfamily
AT juutijarmot identificationandstructuralcharacterizationoflytuauniquepeptidoglycanendopeptidasefromthelysostaphinfamily
AT hiramatsukeiichi identificationandstructuralcharacterizationoflytuauniquepeptidoglycanendopeptidasefromthelysostaphinfamily
AT kontinenvesa identificationandstructuralcharacterizationoflytuauniquepeptidoglycanendopeptidasefromthelysostaphinfamily
AT permiperttu identificationandstructuralcharacterizationoflytuauniquepeptidoglycanendopeptidasefromthelysostaphinfamily