Detection of Lipid-Linked Peptidoglycan Precursors by Exploiting an Unexpected Transpeptidase Reaction.
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| Title: | Detection of Lipid-Linked Peptidoglycan Precursors by Exploiting an Unexpected Transpeptidase Reaction. |
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| Authors: | Yuan Qiao1, Lebar, Matthew D.2, Schirner, Kathrin3, Schaefer, Kaitlin2, Hirokazu Tsukamoto2, Kahne, Daniel1,2 kahne@chemistry.harvard.edu, Walker, Suzanne1,3 suzanne_walker@hms.harvard.edu |
| Source: | Journal of the American Chemical Society. 10/22/2014, Vol. 136 Issue 42, p14678-14681. 4p. 5 Diagrams. |
| Subjects: | Penicillin-binding proteins, Peptidoglycans, Peptidase, Staphylococcus aureus, Chemical synthesis |
| Abstract: | Penicillin-binding proteins (PBPs) are involved in the synthesis and remodeling of bacterial peptidoglycan (PG). Staphylococcus aureus expresses four PBPs. Genetic studies in S. aureus have implicated PBP4 in the formation of highly cross-linked PG, but biochemical studies have not reached a consensus on its primary enzymatic activity. Using synthetic Lipid II, we show here that PBP4 preferentially acts as a transpeptidase (TP) in vitro. Moreover, it is the PBP primarily responsible for incorporating exogenous D-amino acids into cellular PG, implying that it also has TP activity in vivo. Notably, PBP4 efficiently exchanges D-amino acids not only into PG polymers but also into the PG monomers Lipid I and Lipid II. This is the first demonstration that any TP domain of a PBP can activate the PG monomer building blocks. Exploiting the promiscuous TP activity of PBP4, we developed a simple, highly sensitive assay to detect cellular pools of lipid-linked PG precursors, which are of notoriously low abundance. This method, which addresses a longstanding problem, is useful for assessing how genetic and pharmacological perturbations affect precursor levels, and may facilitate studies to elucidate antibiotic mechanism of action. [ABSTRACT FROM AUTHOR] |
| Copyright of Journal of the American Chemical Society is the property of American Chemical Society and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.) | |
| Database: | Engineering Source |
| FullText | Text: Availability: 0 |
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| Header | DbId: egs DbLabel: Engineering Source An: 99392262 AccessLevel: 6 PubType: Academic Journal PubTypeId: academicJournal PreciseRelevancyScore: 0 |
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| Items | – Name: Title Label: Title Group: Ti Data: Detection of Lipid-Linked Peptidoglycan Precursors by Exploiting an Unexpected Transpeptidase Reaction. – Name: Author Label: Authors Group: Au Data: <searchLink fieldCode="AR" term="%22Yuan+Qiao%22">Yuan Qiao</searchLink><relatesTo>1</relatesTo><br /><searchLink fieldCode="AR" term="%22Lebar%2C+Matthew+D%2E%22">Lebar, Matthew D.</searchLink><relatesTo>2</relatesTo><br /><searchLink fieldCode="AR" term="%22Schirner%2C+Kathrin%22">Schirner, Kathrin</searchLink><relatesTo>3</relatesTo><br /><searchLink fieldCode="AR" term="%22Schaefer%2C+Kaitlin%22">Schaefer, Kaitlin</searchLink><relatesTo>2</relatesTo><br /><searchLink fieldCode="AR" term="%22Hirokazu+Tsukamoto%22">Hirokazu Tsukamoto</searchLink><relatesTo>2</relatesTo><br /><searchLink fieldCode="AR" term="%22Kahne%2C+Daniel%22">Kahne, Daniel</searchLink><relatesTo>1,2</relatesTo><i> kahne@chemistry.harvard.edu</i><br /><searchLink fieldCode="AR" term="%22Walker%2C+Suzanne%22">Walker, Suzanne</searchLink><relatesTo>1,3</relatesTo><i> suzanne_walker@hms.harvard.edu</i> – Name: TitleSource Label: Source Group: Src Data: <searchLink fieldCode="JN" term="%22Journal+of+the+American+Chemical+Society%22">Journal of the American Chemical Society</searchLink>. 10/22/2014, Vol. 136 Issue 42, p14678-14681. 4p. 5 Diagrams. – Name: Subject Label: Subjects Group: Su Data: <searchLink fieldCode="DE" term="%22Penicillin-binding+proteins%22">Penicillin-binding proteins</searchLink><br /><searchLink fieldCode="DE" term="%22Peptidoglycans%22">Peptidoglycans</searchLink><br /><searchLink fieldCode="DE" term="%22Peptidase%22">Peptidase</searchLink><br /><searchLink fieldCode="DE" term="%22Staphylococcus+aureus%22">Staphylococcus aureus</searchLink><br /><searchLink fieldCode="DE" term="%22Chemical+synthesis%22">Chemical synthesis</searchLink> – Name: Abstract Label: Abstract Group: Ab Data: Penicillin-binding proteins (PBPs) are involved in the synthesis and remodeling of bacterial peptidoglycan (PG). Staphylococcus aureus expresses four PBPs. Genetic studies in S. aureus have implicated PBP4 in the formation of highly cross-linked PG, but biochemical studies have not reached a consensus on its primary enzymatic activity. Using synthetic Lipid II, we show here that PBP4 preferentially acts as a transpeptidase (TP) in vitro. Moreover, it is the PBP primarily responsible for incorporating exogenous D-amino acids into cellular PG, implying that it also has TP activity in vivo. Notably, PBP4 efficiently exchanges D-amino acids not only into PG polymers but also into the PG monomers Lipid I and Lipid II. This is the first demonstration that any TP domain of a PBP can activate the PG monomer building blocks. Exploiting the promiscuous TP activity of PBP4, we developed a simple, highly sensitive assay to detect cellular pools of lipid-linked PG precursors, which are of notoriously low abundance. This method, which addresses a longstanding problem, is useful for assessing how genetic and pharmacological perturbations affect precursor levels, and may facilitate studies to elucidate antibiotic mechanism of action. [ABSTRACT FROM AUTHOR] – Name: AbstractSuppliedCopyright Label: Group: Ab Data: <i>Copyright of Journal of the American Chemical Society is the property of American Chemical Society and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract.</i> (Copyright applies to all Abstracts.) |
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| RecordInfo | BibRecord: BibEntity: Identifiers: – Type: doi Value: 10.1021/ja508147s Languages: – Code: eng Text: English PhysicalDescription: Pagination: PageCount: 4 StartPage: 14678 Subjects: – SubjectFull: Penicillin-binding proteins Type: general – SubjectFull: Peptidoglycans Type: general – SubjectFull: Peptidase Type: general – SubjectFull: Staphylococcus aureus Type: general – SubjectFull: Chemical synthesis Type: general Titles: – TitleFull: Detection of Lipid-Linked Peptidoglycan Precursors by Exploiting an Unexpected Transpeptidase Reaction. Type: main BibRelationships: HasContributorRelationships: – PersonEntity: Name: NameFull: Yuan Qiao – PersonEntity: Name: NameFull: Lebar, Matthew D. – PersonEntity: Name: NameFull: Schirner, Kathrin – PersonEntity: Name: NameFull: Schaefer, Kaitlin – PersonEntity: Name: NameFull: Hirokazu Tsukamoto – PersonEntity: Name: NameFull: Kahne, Daniel – PersonEntity: Name: NameFull: Walker, Suzanne IsPartOfRelationships: – BibEntity: Dates: – D: 22 M: 10 Text: 10/22/2014 Type: published Y: 2014 Identifiers: – Type: issn-print Value: 00027863 Numbering: – Type: volume Value: 136 – Type: issue Value: 42 Titles: – TitleFull: Journal of the American Chemical Society Type: main |
| ResultId | 1 |
