000205733 001__ 205733
000205733 005__ 20250730145313.0
000205733 0247_ $$2doi$$a10.1128/mBio.01120-13
000205733 0247_ $$2pmid$$apmid:24520066
000205733 0247_ $$2pmc$$apmc:PMC3950521
000205733 0247_ $$2ISSN$$a2150-7511
000205733 0247_ $$2ISSN$$a2161-2129
000205733 0247_ $$2WOS$$aWOS:000332526500038
000205733 0247_ $$2openalex$$aopenalex:W2130883530
000205733 037__ $$aPUBDB-2015-00293
000205733 041__ $$aeng
000205733 082__ $$a570
000205733 1001_ $$0P:(DE-HGF)0$$aMellroth, P.$$b0$$eCorresponding Author
000205733 245__ $$aStructural and Functional Insights Into Peptidoglycan Access for the Lytic Amidase LytA of Streptococcus Pneumoniae
000205733 260__ $$aWashington, DC$$bAmerican Society for Microbiology$$c2014
000205733 3367_ $$2DRIVER$$aarticle
000205733 3367_ $$2DataCite$$aOutput Types/Journal article
000205733 3367_ $$0PUB:(DE-HGF)16$$2PUB:(DE-HGF)$$aJournal Article$$bjournal$$mjournal$$s1426683763_16686
000205733 3367_ $$2BibTeX$$aARTICLE
000205733 3367_ $$2ORCID$$aJOURNAL_ARTICLE
000205733 3367_ $$00$$2EndNote$$aJournal Article
000205733 500__ $$aOA
000205733 520__ $$aThe cytosolic N-acetylmuramoyl-l-alanine amidase LytA protein of Streptococcus pneumoniae, which is released by bacterial lysis, associates with the cell wall via its choline-binding motif. During exponential growth, LytA accesses its peptidoglycan substrate to cause lysis only when nascent peptidoglycan synthesis is stalled by nutrient starvation or β-lactam antibiotics. Here we present three-dimensional structures of LytA and establish the requirements for substrate binding and catalytic activity. The solution structure of the full-length LytA dimer reveals a peculiar fold, with the choline-binding domains forming a rigid V-shaped scaffold and the relatively more flexible amidase domains attached in a trans position. The 1.05-Å crystal structure of the amidase domain reveals a prominent Y-shaped binding crevice composed of three contiguous subregions, with a zinc-containing active site localized at the bottom of the branch point. Site-directed mutagenesis was employed to identify catalytic residues and to investigate the relative impact of potential substrate-interacting residues lining the binding crevice for the lytic activity of LytA. In vitro activity assays using defined muropeptide substrates reveal that LytA utilizes a large substrate recognition interface and requires large muropeptide substrates with several connected saccharides that interact with all subregions of the binding crevice for catalysis. We hypothesize that the substrate requirements restrict LytA to the sites on the cell wall where nascent peptidoglycan synthesis occurs.Streptococcus pneumoniae is a human respiratory tract pathogen responsible for millions of deaths annually. Its major pneumococcal autolysin, LytA, is required for autolysis and fratricidal lysis and functions as a virulence factor that facilitates the spread of toxins and factors involved in immune evasion. LytA is also activated by penicillin and vancomycin and is responsible for the lysis induced by these antibiotics. The factors that regulate the lytic activity of LytA are unclear, but it was recently demonstrated that control is at the level of substrate recognition and that LytA required access to the nascent peptidoglycan. The present study was undertaken to structurally and functionally investigate LytA and its substrate-interacting interface and to determine the requirements for substrate recognition and catalysis. Our results reveal that the amidase domain comprises a complex substrate-binding crevice and needs to interact with a large-motif epitope of peptidoglycan for catalysis.
000205733 536__ $$0G:(DE-H253)POF2-P12-20130405$$aPETRA Beamline P12 (POF2-54G14)$$cPOF2-54G14$$fPOF II$$x0
000205733 588__ $$aDataset connected to CrossRef, bib-pubdb1.desy.de, PubMed,
000205733 650_7 $$2NLM Chemicals$$aMutant Proteins
000205733 650_7 $$2NLM Chemicals$$aPeptidoglycan
000205733 650_7 $$0EC 3.5.1.28$$2NLM Chemicals$$aN-Acetylmuramoyl-L-alanine Amidase
000205733 693__ $$0EXP:(DE-H253)P-P12-20150101$$1EXP:(DE-H253)PETRAIII-20150101$$6EXP:(DE-H253)P-P12-20150101$$aPETRA III$$fPETRA Beamline P12$$x0
000205733 7001_ $$0P:(DE-H253)PIP1017765$$aSandalova, Tatyana$$b1
000205733 7001_ $$0P:(DE-H253)PIP1010068$$aKikhney, Alexey$$b2
000205733 7001_ $$0P:(DE-HGF)0$$aVilaplana, Francisco$$b3
000205733 7001_ $$0P:(DE-HGF)0$$aHesek, Dusan$$b4
000205733 7001_ $$0P:(DE-HGF)0$$aLee, Mijoon$$b5
000205733 7001_ $$0P:(DE-HGF)0$$aMobashery, Shahriar$$b6
000205733 7001_ $$0P:(DE-HGF)0$$aNormark, Staffan$$b7
000205733 7001_ $$0P:(DE-H253)PIP1001422$$aSvergun, Dmitri$$b8
000205733 7001_ $$0P:(DE-HGF)0$$aHenriques-Normark, Birgitta$$b9
000205733 7001_ $$0P:(DE-H253)PIP1018008$$aAchour, Adnane$$b10
000205733 773__ $$0PERI:(DE-600)2557172-2$$a10.1128/mBio.01120-13$$gVol. 5, no. 1, p. e01120-13 - e01120-13$$n1$$pe01120-13$$tmBio$$v5$$x2150-7511$$y2014
000205733 8564_ $$uhttps://bib-pubdb1.desy.de/record/205733/files/mBio-2014-Mellroth-.pdf$$yOpenAccess
000205733 8564_ $$uhttps://bib-pubdb1.desy.de/record/205733/files/mBio-2014-Mellroth-.gif?subformat=icon$$xicon$$yOpenAccess
000205733 8564_ $$uhttps://bib-pubdb1.desy.de/record/205733/files/mBio-2014-Mellroth-.jpg?subformat=icon-180$$xicon-180$$yOpenAccess
000205733 8564_ $$uhttps://bib-pubdb1.desy.de/record/205733/files/mBio-2014-Mellroth-.jpg?subformat=icon-700$$xicon-700$$yOpenAccess
000205733 8564_ $$uhttps://bib-pubdb1.desy.de/record/205733/files/mBio-2014-Mellroth-.pdf?subformat=pdfa$$xpdfa$$yOpenAccess
000205733 8564_ $$uhttps://bib-pubdb1.desy.de/record/205733/files/mBio-2014-Mellroth-.jpg?subformat=icon-1440$$xicon-1440$$yOpenAccess
000205733 8564_ $$uhttps://bib-pubdb1.desy.de/record/205733/files/mBio-2014-Mellroth-.jpg?subformat=icon-640$$xicon-640$$yOpenAccess
000205733 909CO $$ooai:bib-pubdb1.desy.de:205733$$pdnbdelivery$$pdriver$$pVDB$$popen_access$$popenaire
000205733 9101_ $$0I:(DE-HGF)0$$6P:(DE-H253)PIP1017765$$aExternes Institut$$b1$$k>Extern
000205733 9101_ $$0I:(DE-588b)235011-7$$6P:(DE-H253)PIP1010068$$aEuropäisches Laboratorium für Molekularbiologie$$b2$$kEMBL
000205733 9101_ $$0I:(DE-HGF)0$$6P:(DE-H253)PIP1010068$$aExternes Institut$$b2$$k>Extern
000205733 9101_ $$0I:(DE-588b)235011-7$$6P:(DE-H253)PIP1001422$$aEuropäisches Laboratorium für Molekularbiologie$$b8$$kEMBL
000205733 9101_ $$0I:(DE-HGF)0$$6P:(DE-H253)PIP1001422$$aExternes Institut$$b8$$k>Extern
000205733 9101_ $$0I:(DE-HGF)0$$6P:(DE-H253)PIP1018008$$aExternes Institut$$b10$$k>Extern
000205733 9132_ $$0G:(DE-HGF)POF3-622$$1G:(DE-HGF)POF3-620$$2G:(DE-HGF)POF3-600$$9G:(DE-HGF)POF3-6G3$$aDE-HGF$$bForschungsbereich Materie$$lVon Materie zu Materialien und Leben$$vFacility topic: Research on Matter with Brilliant Light Sources$$x0
000205733 9131_ $$0G:(DE-HGF)POF2-54G14$$1G:(DE-HGF)POF2-540$$2G:(DE-HGF)POF2-500$$3G:(DE-HGF)POF2$$4G:(DE-HGF)POF$$9G:(DE-H253)POF2-P12-20130405$$aDE-H253$$bStruktur der Materie$$lForschung mit Photonen, Neutronen, Ionen$$vPETRA III$$x0
000205733 9141_ $$y2014
000205733 915__ $$0StatID:(DE-HGF)0150$$2StatID$$aDBCoverage$$bWeb of Science Core Collection
000205733 915__ $$0StatID:(DE-HGF)1050$$2StatID$$aDBCoverage$$bBIOSIS Previews
000205733 915__ $$0StatID:(DE-HGF)0100$$2StatID$$aJCR
000205733 915__ $$0StatID:(DE-HGF)0500$$2StatID$$aDBCoverage$$bDOAJ
000205733 915__ $$0LIC:(DE-HGF)CCBYNCSA3$$2HGFVOC$$aCreative Commons Attribution-NonCommercial-ShareAlike CC BY-NC-SA 3.0
000205733 915__ $$0StatID:(DE-HGF)0111$$2StatID$$aWoS$$bScience Citation Index Expanded
000205733 915__ $$0StatID:(DE-HGF)0510$$2StatID$$aOpenAccess
000205733 915__ $$0StatID:(DE-HGF)9905$$2StatID$$aIF >= 5
000205733 915__ $$0StatID:(DE-HGF)0310$$2StatID$$aDBCoverage$$bNCBI Molecular Biology Database
000205733 915__ $$0StatID:(DE-HGF)0300$$2StatID$$aDBCoverage$$bMedline
000205733 915__ $$0StatID:(DE-HGF)0199$$2StatID$$aDBCoverage$$bThomson Reuters Master Journal List
000205733 9201_ $$0I:(DE-H253)HAS-User-20120731$$kDOOR$$lDOOR-User$$x0
000205733 980__ $$ajournal
000205733 980__ $$aVDB
000205733 980__ $$aUNRESTRICTED
000205733 980__ $$aFullTexts
000205733 980__ $$aI:(DE-H253)HAS-User-20120731
000205733 9801_ $$aFullTexts