Kinetic mechanism of the Escherichia coli UDPMurNAc-tripeptide D-alanyl-D-alanine-adding enzyme: use of a glutathione S-transferase fusion

Biochemistry
M S AndersonD L Pompliano

Abstract

The D-alanyl-D-alanine-adding enzyme encoded by the murF gene catalyzes the ATP-dependent formation of UDP-N-acetylmuramyl-L-gamma-D-Glu-meso-diaminopimelyl-D-Ala-D-Ala (UDP-MurNAc-tripeptide). MurF has been cloned from Escherichia coli and expressed as a glutathione S-transferase (GST) fusion using the tac promoter-based pGEX-KT vector. From induced, broken cell preparations, highly active fusion was recovered and purified in one step by affinity chromatography. The purified fusion protein was strongly inhibited by substrate UDPMurNAc-tripeptide, a response unaltered by changes in assay pH or by cleavage from the fusion partner. However, this effect was suppressed by the addition of 0.5 M NaCl. Initial velocity and dead-end inhibitor studies with the fusion enzyme were most consistent with a sequential ordered kinetic mechanism for the forward reaction in which ATP binds to free enzyme, followed by tripeptide and D-Ala-D-Ala in sequence prior to product release. Reported homologies between the MurF protein and the three preceding steps of cytoplasmic murein biosynthesis, MurC, -D, and -E, [Ikeda et al. (1990) J. Gen. Appl. Microbiol. 36, 179-187], raise the prospect that all of these enzymes will be found to proceed via this m...Continue Reading

Citations

Oct 28, 2005·Antimicrobial Agents and Chemotherapy·Ellen Z BaumKaren Bush
Dec 27, 2005·Antimicrobial Agents and Chemotherapy·Ellen Z BaumKaren Bush
Oct 3, 2007·Antimicrobial Agents and Chemotherapy·Ellen Z BaumKaren Bush
Dec 23, 2008·BMC Biochemistry·Catherine Paradis-BleauRoger C Levesque
May 28, 2009·Antimicrobial Agents and Chemotherapy·Ellen Z BaumKaren Bush
Feb 10, 2009·Journal of Molecular Modeling·Andrej PerdihTom Solmajer
Jun 15, 2011·Journal of Molecular Modeling·Tihomir TomašićLucija Peterlin Mašič
Dec 21, 2002·Molecular Microbiology·Ahmed El ZoeibyRoger C Levesque
Aug 31, 2013·Journal of Computer-aided Molecular Design·Andrej PerdihTom Solmajer
Nov 28, 2015·Biochimie·Harika VemulaWilliam G Gutheil
Jun 28, 2011·European Journal of Medicinal Chemistry·Tihomir TomašićLucija Peterlin Mašič
Sep 4, 2010·Computer Methods and Programs in Biomedicine·J G HattersleyN D Evans
Nov 19, 2008·Bioorganic & Medicinal Chemistry Letters·Tihomir TomasićLucija Peterlin Masic
Dec 3, 2005·Protein Science : a Publication of the Protein Society·Kenton L LongeneckerVincent S Stoll
Feb 24, 2006·Chemical Biology & Drug Design·Geoffrey F StamperVincent S Stoll
Aug 19, 2014·Molecular Microbiology·Imène KouidmiCatherine Paradis-Bleau
Dec 22, 2009·ChemMedChem·Tihomir TomasićLucija Peterlin Masic
Aug 29, 2006·Journal of Molecular Biology·Clyde A Smith
Nov 20, 2015·Scientific Reports·Tomohide SaioFuyuhiko Inagaki
Apr 30, 2009·The Biochemical Journal·Catherine Paradis-BleauRoger C Levesque
Mar 21, 2002·Expert Opinion on Therapeutic Targets·David W Green
Feb 13, 2008·FEMS Microbiology Reviews·Hélène BarreteauDidier Blanot
Jul 1, 2014·Biochemical and Biophysical Research Communications·Sun-Shin ChaKyung Min Chung
Aug 9, 2012·ACS Medicinal Chemistry Letters·Tihomir TomašićLucija Peterlin Mašič
Apr 15, 2014·Journal of Chemical Information and Modeling·Andrej PerdihTom Solmajer
Apr 21, 2017·Nature Communications·Hai LiangCatherine L Grimes
Jun 12, 2019·FEMS Microbiology Reviews·Arundhati MaitraSanjib Bhakta
Nov 26, 2010·Critical Reviews in Biotechnology·Ankur GautamRupinder Tewari
Apr 1, 2005·Journal of Biomolecular Screening·S M SolapureS M De Sousa
Dec 2, 2014·Biomolecular Concepts·Roman ŠinkDidier Blanot
Aug 5, 2017·Journal of Receptor and Signal Transduction Research·Mohammed Afzal Azam, Srikanth Jupudi
Aug 11, 2004·The Journal of Biological Chemistry·Joseph M Jez, Rebecca E Cahoon

❮ Previous
Next ❯

Related Concepts

Related Feeds

Bacterial Cell Wall Structure (ASM)

Bacterial cell walls are made of peptidoglycan (also called murein), which is made from polysaccharide chains cross-linked by unusual peptides containing D-amino acids. Here is the latest research on bacterial cell wall structures.

Bacterial Cell Wall Structure

Bacterial cell walls are made of peptidoglycan (also called murein), which is made from polysaccharide chains cross-linked by unusual peptides containing D-amino acids. Here is the latest research on bacterial cell wall structures.