| IED ID | IndEnz0002001141 |
| Enzyme Type ID | protease001141 |
| Protein Name |
Penicillin-binding protein 2a PBP2a Cell wall synthase PBP2a Includes: Penicillin-insensitive transglycosylase EC 2.4.1.129 Peptidoglycan TGase Peptidoglycan glycosyltransferase ; Penicillin-sensitive transpeptidase EC 3.4.16.4 DD-transpeptidase |
| Gene Name | pbp2a spr1823 |
| Organism | Streptococcus pneumoniae (strain ATCC BAA-255 / R6) |
| Taxonomic Lineage | cellular organisms Bacteria Terrabacteria group Firmicutes Bacilli Lactobacillales Streptococcaceae Streptococcus Streptococcus pneumoniae Streptococcus pneumoniae (strain ATCC BAA-255 / R6) |
| Enzyme Sequence | MKLDKLFEKFLSLFKKETSELEDSDSTILRRSRSDRKKLAQVGPIRKFWRRYHLTKIILILGLSAGLLVGIYLFAVAKSTNVNDLQNALKTRTLIFDREEKEAGALSGQKGTYVELTDISKNLQNAVIATEDRSFYKNDGINYGRFFLAIVTAGRSGGGSTITQQLAKNAYLSQDQTVERKAKEFFLALELSKKYSKEQILTMYLNNAYFGNGVWGVEDASKKYFGVSASEVSLDQAATLAGMLKGPELYNPLNSVEDSTNRRDTVLQNMVAAGYIDKNQETEAAEVDMTSQLHDKYEGKISDYRYPSYFDAVVNEAVSKYNLTEEEIVNNGYRIYTELDQNYQANMQIVYENTSLFPRAEDGTFAQSGSVALEPKTGGVRGVVGQVADNDKTGFRNFNYATQSKRSPGSTIKPLVVYTPAVEAGWALNKQLDNHTMQYDSYKVDNYAGIKTSREVPMYQSLAESLNLPAVATVNDLGVDKAFEAGEKFGLNMEKVDRVLGVALGSGVETNPLQMAQAYAAFANEGLMPEAHFISRIENASGQVIASHKNSQKRVIDKSVADKMTSMMLGTFTNGTGISSSPADYVMAGKTGTTEAVFNPEYTSDQWVIGYTPDVVISHWLGFPTTDENHYLAGSTSNGAAHVFRNIANTILPYTPGSTFTVENAYKQNGIAPANTKRQVQTNDNSQTDDNLSDIRGRAQSLVDEASRAISDAKIKEKAQTIWDSIVNLFR |
| Enzyme Length | 731 |
| Uniprot Accession Number | Q8DNB6 |
| Absorption | |
| Active Site | ACT_SITE 131; /note=Proton donor; for transglycosylase activity; /evidence=ECO:0000269|PubMed:22487093; ACT_SITE 410; /note=Acyl-ester intermediate; for transpeptidase activity; /evidence=ECO:0000250|UniProtKB:P02919 |
| Activity Regulation | ACTIVITY REGULATION: Optimal transglycosylase/glycosyltransferase (GT) activity in the presence of 30-40% dimethylsulfoxide and 0.002% Triton X-100. High GT activity in the presence of CHAPS, Triton X-100 and n-dodecyl-beta-D-maltopyranoside (DDM) detergents, and to a lesser extent in the presence of Cymal-5 (PubMed:22487093). GT activity is inhibited by moenomycin (PubMed:12867450, PubMed:22487093). 50% inhibition of the GT activity with 2.8 uM moenomycin. No effect on GT activity detected with 2.8 uM vancomycin, but complete inhibition with 28 uM vancomycin (PubMed:12867450). No GT activity in the presence of n-octyl-beta-D-glucopyranoside, Cymal-3 and Cymal-4 detergents (PubMed:22487093). {ECO:0000269|PubMed:12867450, ECO:0000269|PubMed:22487093}. |
| Binding Site | |
| Calcium Binding | |
| catalytic Activity | CATALYTIC ACTIVITY: Reaction=Preferential cleavage: (Ac)(2)-L-Lys-D-Ala-|-D-Ala. Also transpeptidation of peptidyl-alanyl moieties that are N-acyl substituents of D-alanine.; EC=3.4.16.4; Evidence={ECO:0000269|PubMed:10217767, ECO:0000269|PubMed:22487093}; CATALYTIC ACTIVITY: Reaction=[GlcNAc-(1->4)-Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)](n)-di-trans,octa-cis-undecaprenyl diphosphate + beta-D-GlcNAc-(1->4)-Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-di-trans,octa-cis-undecaprenyl diphosphate = [GlcNAc-(1->4)-Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)](n+1)-di-trans-octa-cis-undecaprenyl diphosphate + di-trans,octa-cis-undecaprenyl diphosphate + H(+); Xref=Rhea:RHEA:23708, Rhea:RHEA-COMP:9602, Rhea:RHEA-COMP:9603, ChEBI:CHEBI:15378, ChEBI:CHEBI:58405, ChEBI:CHEBI:60033, ChEBI:CHEBI:78435; EC=2.4.1.129; Evidence={ECO:0000269|PubMed:12867450, ECO:0000269|PubMed:22487093}; |
| DNA Binding | |
| EC Number | 2.4.1.129; 3.4.16.4 |
| Enzyme Function | FUNCTION: Cell wall formation. Synthesis of cross-linked peptidoglycan (PG) from the lipid intermediates (By similarity). Binds dansylated lipid II and catalyzes the polymerization of glycan chains (PubMed:12867450, PubMed:22487093). Hydrolyzes S2d (N-benzoyl-D-alanylmercaptoacetic acid) molecule, a synthetic thiolester analog of cell wall stem peptide (PubMed:10217767, PubMed:22487093). Active against bocillin, a fluorescent penicillin. No transpeptidase activity with non-fluorescent lysine-containing lipid II as substrate (PubMed:22487093). {ECO:0000250|UniProtKB:P02918, ECO:0000269|PubMed:10217767, ECO:0000269|PubMed:12867450, ECO:0000269|PubMed:22487093}. |
| Temperature Dependency | |
| PH Dependency | |
| Pathway | PATHWAY: Cell wall biogenesis; peptidoglycan biosynthesis. {ECO:0000305}. |
| nucleotide Binding | |
| Features | Active site (2); Chain (1); Mutagenesis (1); Region (4); Sequence conflict (2); Topological domain (2); Transmembrane (1) |
| Keywords | Acyltransferase;Antibiotic resistance;Carboxypeptidase;Cell membrane;Cell shape;Cell wall;Cell wall biogenesis/degradation;Direct protein sequencing;Glycosyltransferase;Hydrolase;Membrane;Multifunctional enzyme;Peptidoglycan synthesis;Protease;Reference proteome;Secreted;Signal-anchor;Transferase;Transmembrane;Transmembrane helix |
| Interact With | |
| Induction | |
| Subcellular Location | SUBCELLULAR LOCATION: Cell membrane {ECO:0000269|PubMed:10217767, ECO:0000269|PubMed:12867450, ECO:0000269|PubMed:22487093}; Single-pass type II membrane protein {ECO:0000269|PubMed:10217767, ECO:0000269|PubMed:12867450, ECO:0000269|PubMed:22487093}. Secreted, cell wall {ECO:0000305}. Note=Localizes to sites of new peptidoglycan (PG) synthesis at midcell independently of MacP. {ECO:0000250|UniProtKB:A0A0H2ZMF9}. |
| Modified Residue | |
| Post Translational Modification | |
| Signal Peptide | |
| Structure 3D | |
| Cross Reference PDB | - |
| Mapped Pubmed ID | - |
| Motif | |
| Gene Encoded By | |
| Mass | 80,799 |
| Kinetics | BIOPHYSICOCHEMICAL PROPERTIES: Kinetic parameters: KM=40.6 uM for dansylated lipid II {ECO:0000269|PubMed:12867450}; Vmax=0.38 nmol/min/mg enzyme with lysine-dansylated lipid II as substrate (at pH 7.5 and 30 degrees Celsius in the presence of 25% dimethylsulfoxide and 0.04 Triton X-100) {ECO:0000269|PubMed:22487093}; Vmax=403 nmol/min/mg enzyme for the hydrolysis of S2d (N-benzoyl-D-alanylmercaptoacetic acid) (at pH 7.0 and 37 degrees Celsius) {ECO:0000269|PubMed:22487093}; Note=kcat is greater than 0.0005 sec(-1) with dansylated lipid II as substrate (PubMed:22487093). kcat/KM is 0.001 M(-1)sec(-1) with dansylated lipid II as substrate (PubMed:12867450). {ECO:0000269|PubMed:12867450, ECO:0000269|PubMed:22487093}; |
| Metal Binding | |
| Rhea ID | RHEA:23708 |
| Cross Reference Brenda |