title: Crystal Structure of the N-Acetylmuramic Acid α-1-Phosphate (MurNAc-α1-P) Uridylyltransferase MurU, a Minimal Sugar Nucleotidyltransferase and Potential Drug Target Enzyme in Gram-negative Pathogens.
abstract: The N-acetylmuramic acid α-1-phosphate (MurNAc-α1-P) uridylyltransferase MurU catalyzes the synthesis of uridine diphosphate (UDP)-MurNAc, a crucial precursor of the bacterial peptidoglycan cell wall. MurU is part of a recently identified cell wall recycling pathway in Gram-negative bacteria that bypasses the general de novo biosynthesis of UDP-MurNAc and contributes to high intrinsic resistance to the antibiotic fosfomycin, which targets UDP-MurNAc de novo biosynthesis. To provide insights into substrate binding and specificity, we solved crystal structures of MurU of Pseudomonas putida in native and ligand-bound states at high resolution. With the help of these structures, critical enzyme-substrate interactions were identified that enable tight binding of MurNAc-α1-P to the active site of MurU. The MurU structures define a "minimal domain" required for general nucleotidyltransferase activity. They furthermore provide a structural basis for the chemical design of inhibitors of MurU that could serve as novel drugs in combination therapy against multidrug-resistant Gram-negative pathogens.
authors.affiliation: From the Interfaculty Institute of Biochemistry (IFIB).
authors.forename: Michaela
authors.initials: M
authors.lastname: Renner-Schneck
authors.affiliation: Interfaculty Institute of Microbiology and Infection Medicine Tübingen (IMIT), Department of Biology, and.
authors.forename: Isabel
authors.initials: I
authors.lastname: Hinderberger
authors.affiliation: Interfaculty Institute of Microbiology and Infection Medicine Tübingen (IMIT), Department of Biology, and.
authors.forename: Jonathan
authors.initials: J
authors.lastname: Gisin
authors.affiliation: Institute of Pharmacy, University of Tübingen, 72076 Tübingen, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Exner
authors.affiliation: Interfaculty Institute of Microbiology and Infection Medicine Tübingen (IMIT), Department of Biology, and christoph.mayer@uni-tuebingen.de.
authors.forename: Christoph
authors.initials: C
authors.lastname: Mayer
authors.affiliation: From the Interfaculty Institute of Biochemistry (IFIB), thilo.stehle@uni-tuebingen.de.
authors.forename: Thilo
authors.initials: T
authors.lastname: Stehle
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D020134:Catalytic Domain; D003001:Cloning, Molecular; D018360:Crystallography, X-Ray; D006090:Gram-Negative Bacteria / Q000187:drug effects / Q000201:enzymology / Q000472:pathogenicity; D008274:Magnesium / Q000737:chemistry; D008958:Models, Molecular; D009713:Nucleotidyltransferases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D016958:Pseudomonas putida / Q000201:enzymology / Q000235:genetics; D013379:Substrate Specificity; D014538:Uridine Diphosphate N-Acetylmuramic Acid / Q000096:biosynthesis
chemical_list: D001426:Bacterial Proteins; D014538:Uridine Diphosphate N-Acetylmuramic Acid; D009713:Nucleotidyltransferases; D008274:Magnesium
keywords: Bacteria; Cell Wall; Drug Resistance; Enzyme Structure; Fosfomycin; N-Acetylmuramic Acid α-1-Phosphate; Peptidoglycan Biosynthesis; Structural Biology; UDP-MurNAc