Introduction
(TP), also known as penicillin-binding proteins (PBP),
catalyze the cross-linking of peptidoglycan polymers during bacterial cell wall
synthesis. The natural transpeptidase substrate is the D-Ala-D-Ala
peptidoglycan side chain terminus. Beta-lactam (β-lactam) antibiotics, which
include penicillins, cephalosporins and carbapenems, bind and irreversibly
inhibit transpeptidases by mimicking the D-Ala-D-Ala substrate, resulting in
the inhibition of cell wall synthesis and ultimately bacterial cell growth.
Overuse and misuse of β-lactams has led to the generation of methicillin resistant
Staphylococcus aureus (MRSA) isolates that have acquired an
alternative transpeptidase, PBP2a, which is neither bound nor inhibited by β-
lactams. MRSA isolates are resistant to all β-lactams, can be hospital- or
community-acquired, and are often the cause of significant morbidity and
mortality. Furthermore, they are often only susceptible to “last resort
antibiotics”, such as vancomycin. Recently, two cephalosporins - ceftobiprole
and ceftaroline - that bind and inhibit PBP2a have been developed.
β-lactam antibiotics, which include penicillins, cephalosporins and carbapenems,
have been used to treat Staphylococcus aureus infections. The overuse and misuse
of β-lactam antibiotics has led to strains of Staphylococcus aureus that are
resistant to all β-lactams; so called MRSA strains. MRSA can be hospital- or
community-acquired and are often the cause of significant morbidity and mortality.
β-Lactam antibiotics stop the production of the cell wall by targeting bacterial
PBPs. The cell wall, which is composed of peptidoglycan and surrounds the cell
membrane, is crucial for maintaining the structural integrity of the bacterium.
The cell wall is composed of rows of peptidoglycan cross-linked together with
pentaglycine chains. Peptidoglycan consists of N-acetylmuramic Acid (NAM) and
N-acetylglucosamine (NAG) polymers. The NAM residues have a five amino acid side
chain that terminates with two D-Alanine (D-Ala) residues. MRSA is resistant to all
β-lactams because it acquires an alternative PBP, PBP2a, that is not bound or
inhibited by any β-lactams. Recently, two cephalosporins - ceftobiprole and
ceftaroline - that bind and inhibit PBP2a have been developed.
How does PBP2a works?
PBP2a is composed of two domains: a (NPB) domain and a
domain. The NBP domain of PBP2a is anchored in the cell membrane, while the TP domain “sits” in the periplasm with its active site facing the inner surface of the cell wall. The active site contains a
serine residue at position 403 () which catalyzes the cross-linking of the peptidoglycan rows with pentaglycine cross-links.
Catalytic Mechanism of PBP2a
The D-Ala-D-Ala side-chain substrate of the peptidoglycan accesses the active site of the PBP2a.
Ser403 nucleophilically attacks the peptide bond of the terminal D-Ala residues of the substrate. The terminal D-Ala residue then exits the active site.
The now terminal D-Ala residue forms a covalent bond to Ser403, while a cross-linking pentaglycine chain enters the active site.
A covalent bond forms between the pentaglycine chain and the terminal D-Ala residue, regenerating the active site serine residue.
