The structure of the N-terminal module of the cell wall hydrolase RipA and its role in regulating catalytic activity
(2018) In Proteins: Structure, Function and Bioinformatics 86(9). p.912-923- Abstract
RipA plays a vital role during cell division of Mycobacterium tuberculosis by degrading the cell wall peptidoglycan at the septum, allowing daughter cell separation. The peptidoglycan degrading activity relies on the NlpC/P60 domain, and as it is potentially harmful when deregulated, spatial and temporal control is necessary in this process. The N-terminal domain of RipA has been proposed to play an inhibitory role blocking the C-terminal NlpC/P60 domain. Accessibility of the active site cysteine residue is however not limited by the presence of the N-terminal domain, but by the lid-module of the inter-domain linker, which is situated in the peptide binding groove of the crystal structures of the catalytic domain. The 2.2 Å resolution... (More)
RipA plays a vital role during cell division of Mycobacterium tuberculosis by degrading the cell wall peptidoglycan at the septum, allowing daughter cell separation. The peptidoglycan degrading activity relies on the NlpC/P60 domain, and as it is potentially harmful when deregulated, spatial and temporal control is necessary in this process. The N-terminal domain of RipA has been proposed to play an inhibitory role blocking the C-terminal NlpC/P60 domain. Accessibility of the active site cysteine residue is however not limited by the presence of the N-terminal domain, but by the lid-module of the inter-domain linker, which is situated in the peptide binding groove of the crystal structures of the catalytic domain. The 2.2 Å resolution structure of the N-terminal domain, determined by Se-SAD phasing, reveals an all-α-fold with 2 long α-helices, and shows similarity to bacterial periplasmic protein domains with scaffold-building role. Size exclusion chromatography and SAXS experiments are consistent with dimer formation of this domain in solution. The SAXS data from the periplasmic two-domain RipA construct suggest a rigid baton-like structure of the N-terminal module, with the catalytic domain connected by a 24 residue long flexible linker. This flexible linker allows for a catalytic zone, which is part of the spatiotemporal control of peptidoglycan degradation.
(Less)
- author
- Steiner, Eva Maria
LU
; Lyngsø, Jeppe ; Guy, Jodie E. ; Bourenkov, Gleb ; Lindqvist, Ylva ; Schneider, Thomas R. ; Pedersen, Jan Skov ; Schneider, Gunter and Schnell, Robert
- publishing date
- 2018-09
- type
- Contribution to journal
- publication status
- published
- keywords
- cell wall remodeling, NlpC/P60, peptidoglycan cleavage, protein structure, SAXS, X-ray crystallography
- in
- Proteins: Structure, Function and Bioinformatics
- volume
- 86
- issue
- 9
- pages
- 912 - 923
- publisher
- John Wiley & Sons Inc.
- external identifiers
-
- pmid:29722065
- scopus:85047815945
- ISSN
- 0887-3585
- DOI
- 10.1002/prot.25523
- language
- English
- LU publication?
- no
- additional info
- Publisher Copyright: © 2018 Wiley Periodicals, Inc.
- id
- 643e4c13-9215-4214-8fc0-04bc5d9adb62
- date added to LUP
- 2024-06-24 11:30:12
- date last changed
- 2024-06-25 03:04:13
@article{643e4c13-9215-4214-8fc0-04bc5d9adb62, abstract = {{<p>RipA plays a vital role during cell division of Mycobacterium tuberculosis by degrading the cell wall peptidoglycan at the septum, allowing daughter cell separation. The peptidoglycan degrading activity relies on the NlpC/P60 domain, and as it is potentially harmful when deregulated, spatial and temporal control is necessary in this process. The N-terminal domain of RipA has been proposed to play an inhibitory role blocking the C-terminal NlpC/P60 domain. Accessibility of the active site cysteine residue is however not limited by the presence of the N-terminal domain, but by the lid-module of the inter-domain linker, which is situated in the peptide binding groove of the crystal structures of the catalytic domain. The 2.2 Å resolution structure of the N-terminal domain, determined by Se-SAD phasing, reveals an all-α-fold with 2 long α-helices, and shows similarity to bacterial periplasmic protein domains with scaffold-building role. Size exclusion chromatography and SAXS experiments are consistent with dimer formation of this domain in solution. The SAXS data from the periplasmic two-domain RipA construct suggest a rigid baton-like structure of the N-terminal module, with the catalytic domain connected by a 24 residue long flexible linker. This flexible linker allows for a catalytic zone, which is part of the spatiotemporal control of peptidoglycan degradation.</p>}}, author = {{Steiner, Eva Maria and Lyngsø, Jeppe and Guy, Jodie E. and Bourenkov, Gleb and Lindqvist, Ylva and Schneider, Thomas R. and Pedersen, Jan Skov and Schneider, Gunter and Schnell, Robert}}, issn = {{0887-3585}}, keywords = {{cell wall remodeling; NlpC/P60; peptidoglycan cleavage; protein structure; SAXS; X-ray crystallography}}, language = {{eng}}, number = {{9}}, pages = {{912--923}}, publisher = {{John Wiley & Sons Inc.}}, series = {{Proteins: Structure, Function and Bioinformatics}}, title = {{The structure of the N-terminal module of the cell wall hydrolase RipA and its role in regulating catalytic activity}}, url = {{http://dx.doi.org/10.1002/prot.25523}}, doi = {{10.1002/prot.25523}}, volume = {{86}}, year = {{2018}}, }