Mechanisms of antimicrobial

action directed against the

bacterial cell wall and
corresponding resistance
mechanisms
M-4 Advanced
Therapeutics
Course

Mechanisms of antimicrobial resistance

Drug-modifying
enzymes

(e.g., - lactamases,
aminoglycosidemodifying enzymes)

Altered
drug
targets
(e.g., PBPs
ribosomes,
DNA gyrase)

Altered
uptake or
accumulation of
drug
(e.g., altered porins,
membrane
efflux pumps)

Subunits for cell wall construction

N-acetylglucosamine

N-acetylmuramic acid

pentapeptide

D-ala-D-ala

Cell Wall Assembly

Second layer of cell

wall cross-linked to
the lower layer

Layer of cell wall

with cross links
of 5 glycines
(gray)

A subunit is added
to the growing chain
Transpeptidase (PBP) forms a 5-glycine bridge between peptides

Transpeptidase, or PBP
(orange sunburst)
is bound by beta-lactam antibiotic (light blue)
and its activity is inhibited
(turns gray)

5-glycine crosslinking bridges cannot form in the presence of a

beta-lactam, and the cell wall is deformed and weakened

Mechanisms of beta-lactam resistance

Drug-modifying enzymes (beta-lactamases)

Gram-positives(e.g., S. aureus) excrete the enzyme

Gram-negative (e.g., E. coli) retain the enzyme in the
periplasm

Overexpression of

cell wall synthetic enzymes

e.g., vancomycin-intermediate S. aureus (VISA)

Alteration of the PBPs so antibiotic cannot bind

e.g., S. pneumoniae, gonococcus

Exclusion from the site of cell wall synthesis

Porin mutations in the outer membrane of Gramnegative bacteria only (e.g., Ps. aeruginosa)

Beta-lactamases
Beta-lactamases

(dark orange)

bind to the antibiotics

(light blue)
and cleave the beta-lactam ring.
The antibiotic is no longer able to inhibit the
function of PBP
(orange sunburst)

Beta-lactamase activity

Altered drug targets

Vancomycin-intermediate S. aureus
Production of excessive cell wall; the antibiotic cannot keep up

MRSA
vancomycin MIC = 2 g/ml

VISA
vancomycin MIC =8 g/ml

MRSA

VISA

Mechanism of vancomycin action

V
D-ala-D-ala

Mechanism of vancomycin resistance

Vancomycin is
unable to bind
to the D-ala-Dlactate structure
D-ala-D-lactate

June 2002: isolated from the catheter exit site in a chronic

dialysis patient
The patient had received multiple courses of abx since
April 2001; toe amputation in April 2002 --> MRSA
bacteremia
VRSA also found at amputation stump wound (with VRE
and Klebsiella); not in the patients nose
Vancomycin MIC >128mcg/ml!! (contains vanA)
Sensitive to trim/sulfa, chloro, tetracyclines, Synercid,
linezolid

MRSA and penicillin-resistant S.

pneumoniae
These bacteria are both resistant because they have

altered bacterial targets -- penicillin-binding proteins

(PBPs or transpeptidases)
In MRSA, the altered PBP2 (mecA) gene is acquired by
gene transfer from another bacterium.
In pneumococci, the alteration in PBP is generated by
uptake of DNA released by dead oral streptococci and
recombination at the pneumococcal pbp gene to create a
new, chimeric protein that does not bind penicillin.

depicted on the next slide . . .

DNA

alpha-strep pbp

Alpha-strep
S. pneumoniae

transformation

alpha-strep pbp

S. pneumoniae chromosomal pbp; penicillin-sensitive

Chimeric pbp (resistant to penicillin)

Outer membrane permeability in

Gram-negative bacteria
Beta-lactam (blue)
enters through an outer
membrane porin channel

Altered porin channel

prevents access of the
antibiotic to the cell wall

Outer membrane
Cell wall
(peptidoglycan)
Inner membrane
Cytoplasm

Bacterium