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Literature summary extracted from

  • Egan, A.J.; Vollmer, W.
    Continuous fluorescence assay for peptidoglycan glycosyltransferases (2016), Methods Mol. Biol., 1440, 171-184 .
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
2.4.1.129 DMSO 20% DMSO has a positive effect on the GTase rate of PBP1B, particularly at the higher Triton X-100 concentration of 0.2%. At higher Triton X-100 concentration in the presence of 20% DMSO the stimulation of PBP1B by LpoB is only 1.6fold Escherichia coli

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.4.1.129 overexpression of His-tagged enzyme in Escherichia coli strain BL21 pDML924 Escherichia coli

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.4.1.129 additional information inhibitory effect of high detergent concentration on the enzyme activity. 25% Dimethylsulfoxide (DMSO) abrogates this detergent effect Escherichia coli

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
2.4.1.129 membrane
-
 Escherichia coli 16020
-

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.4.1.129 Mg2+ required Escherichia coli

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.4.1.129 additional information Escherichia coli natural substrate is lipid II ?
-
-
2.4.1.129 [GlcNAc-(1->4)-Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)]n-diphosphoundecaprenol + GlcNAc-(1->4)-Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphosphoundecaprenol Escherichia coli
-
 [GlcNAc-(1->4)-Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)]n+1-diphosphoundecaprenol + undecaprenyl diphosphate
-
 ?

Organic Solvent Stability

EC Number Organic Solvent Comment Organism
2.4.1.129 additional information organic solvent DMSO is a component of in vitro peptidoglycan synthesis assays making use of its ability to solubilize the lipid II substrate without denaturing enzymes. DMSO at high concentration can potentially affect protein stability and protein-protein interactions. The presence of 20% DMSO severely impairs the interaction of PBP1B with LpoB. In contrast, at lower concentration of Triton X-100 (between 0.02% and 0.08%) the activity of PBP1B can be robustly assayed and its interactions with LpoB, CpoB, TolA and FtsN are maintained. Inhibitory effect of high detergent concentration on the enzyme activity. 25% Dimethylsulfoxide (DMSO) abrogates this detergent effect Escherichia coli

Organism

EC Number Organism UniProt Comment Textmining
2.4.1.129 Escherichia coli P02919
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.4.1.129 gene mrcB, recombinant His-tagged PBP1B from Escherichia coli strain BL21 pDML924 membrane fraction by ultracentrifugation, nickel affinity chromatography, dialysis, and tag cleavage by thrombin, followed by dialysis, cation exchange chromatography and again diaylsis, method overview Escherichia coli

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.4.1.129 additional information natural substrate is lipid II Escherichia coli ?
-
-
2.4.1.129 additional information fluorescence enzyme assay optimization and evaluation, detailed overview Escherichia coli ?
-
-
2.4.1.129 [GlcNAc-(1->4)-Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)]n-diphosphoundecaprenol + GlcNAc-(1->4)-Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphosphoundecaprenol
-
 Escherichia coli [GlcNAc-(1->4)-Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)]n+1-diphosphoundecaprenol + undecaprenyl diphosphate
-
 ?

Synonyms

EC Number Synonyms Comment Organism
2.4.1.129 DD-transpeptidase
-
 Escherichia coli
2.4.1.129 GTase
-
 Escherichia coli
2.4.1.129 PBP
-
 Escherichia coli
2.4.1.129 PBP1b
-
 Escherichia coli
2.4.1.129 penicillin-binding protein
-
 Escherichia coli

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.4.1.129 22
-
 assay at room temperature Escherichia coli

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.4.1.129 7.5
-
 assay at Escherichia coli

General Information

EC Number General Information Comment Organism
2.4.1.129 physiological function the majority of bacteria surround their cytoplasmic membrane with a peptidoglycan sacculus, a continuous layer that is required to maintain cell shape and osmotic stability. The basic chemical structure of peptidoglycan is well known, glycan strands consisting of alternating N -acetylglucosamine (GlcNAc) and N-acetylmuramic acid (MurNAc) residues connected by short stem peptides protruding from MurNAc. Peptides of neighboring glycan strands may be connected (i.e. cross-linked) forming a net-like layer. Peptide cross-links are formed by DD-transpeptidases. DD-transpeptidases covalently bind beta-lactam antibiotics such as penicillin, and are hence named penicillin-binding proteins (PBPs). Most bacteria possess several peptidoglycan synthases capable of catalyzing the glycosyltransferase (GTase) and/or transpeptidase (TPase) reactions. The Gram negative model organism Escherichia coli has three enzymes capable of performing both reactions, so-called bifunctional synthases: PBP1A, PBP1B, and PBP1C, two monofunctional transpeptidases, PBP2 and PBP3, and the monofunctional glycosyltransferase MtgA. The main peptidoglycan synthesis activity in the cell is provided by the semi-redundant PBP1A and PBP1B in consort with the transpeptidases PBP2 and PBP3, latter have essential roles in cell elongation and division, respectively. PBP1C and MtgA are dispensable for growth Escherichia coli