English   español  
Please use this identifier to cite or link to this item: http://hdl.handle.net/10261/177975
Share/Impact:
Statistics
logo share SHARE logo core CORE   Add this article to your Mendeley library MendeleyBASE

Visualizar otros formatos: MARC | Dublin Core | RDF | ORE | MODS | METS | DIDL
Exportar a otros formatos:
Title

A Structural Dissection of the Active Site of the Lytic Transglycosylase MltE from Escherichia coli

AuthorsDik, D.A.; Dik, D.A.; Batuecas, M.T.; Lee, M.; Mahasenan, K.V.; Marous, D.R.; Lastochkin, E.; Fisher, J.F.; Hermoso, J.A.; Mobashery, S.; Lee, M.; Mahasenan, K.V.; Marous, D.R.; Lastochkin, E.; Fisher, J.F.; Hermoso, Juan A; Mobashery, S.
Issue Date2018
PublisherAmerican Chemical Society
CitationBiochemistry 57: 6090- 6098 (2018)
AbstractLytic transglycosylases (LTs) are bacterial enzymes that catalyze the cleavage of the glycan strands of the bacterial cell wall. The mechanism of this cleavage is a remarkable intramolecular transacetalization reaction, accomplished by an ensemble of active-site residues. Because the LT reaction occurs in parallel with the cell wall bond-forming reactions catalyzed by the penicillin-binding proteins, simultaneous inhibition of both enzymes can be particularly bactericidal to Gram-negative bacteria. The MltE lytic transglycosylase is the smallest of the eight LTs encoded by the Escherichia coli genome. Prior crystallographic and computational studies identified four active-site residues - E64, S73, S75, and Y192 - as playing roles in catalysis. Each of these four residues was individually altered by mutation to give four variant enzymes (E64Q, S73A, S75A, and Y192F). All four variants showed reduced catalytic activity [soluble wild type (100%) > soluble Y192F and S75A (both 40%) > S73A (4%) > E64Q (≤1%)]. The crystal structure of each variant protein was determined at the resolution of 2.12 Å for E64Q, 2.33 Å for Y192F, 1.38 Å for S73A, and 1.35 Å for S75A. These variants show alteration of the hydrogen-bond interactions of the active site. Within the framework of a prior computational study of the LT mechanism, we suggest the mechanistic role of these four active-site residues in MltE catalysis.
URIhttp://hdl.handle.net/10261/177975
Identifiersdoi: 10.1021/acs.biochem.8b00800
issn: 1520-4995
Appears in Collections:(IQFR) Artículos
Files in This Item:
File Description SizeFormat 
accesoRestringido.pdf15,38 kBAdobe PDFThumbnail
View/Open
Show full item record
 

Related articles:


WARNING: Items in Digital.CSIC are protected by copyright, with all rights reserved, unless otherwise indicated.