Transpeptidase inhibitors

Dringen, R., Kranich, O., and Hamprecht, B. (1997). The y-glutamyl transpeptidase inhibitor adv-icin preserves glutathione released by astroglial cells in culture. Neurochem. Res. 22 727-733. 

Hilgier, W., Puka, M., and Albrecht, J. 1992. Characteristics of large neutral amino acid-induced release of preloaded 1-glutamine from rat cerebral capillaries in vitro effects of ammonia, hepatic encephalopathy, and gamma-glutamyl transpeptidase inhibitors. J. Neurosci. Res. 32 221-226. 

The answer is d. (Hardman, pp 1097—1098.) The antibiotic clavu-lanic acid is a potent inhibitor of p-lactamases. The mode of inhibition is irreversible. Although clavulanic acid does not effectively inhibit the transpeptidase, it maybe used in conjunction with a p-lactamase-sensitive penicillin to potentiate its activity... 

A term used to describe inhibition induced by a second inhibitor. A particular inhibitor (F) may be a weak inhibitor for a certain enzyme however, in the presence of a second inhibitor (F), inhibition is greatly enhanced. An example of such a system is the inhibitory effect of inorganic pyrophosphate on trichodiene synthase by certain aza analogues Both enantiomers of serine are weak inhibitors of y-glutamyl transpeptidase (L-serine has a Ki value of 10.7 mM), but in the presence of borate ion, they are potent inhibitors ... 

Is specific for Gram-positive microorganisms Is a potent inhibitor of cell-wall transpeptidase Inactivates bacterial P-lactamases Has a spectrum of activity similar to that of penicillin G... 

Several classes of (3-lactamases, often encoded in transmissible plasmids, have spread worldwide rapidly among bacteria, seriously decreasing the effectivenss of penicillins and other (3-lactam anti-biotics.t y Most (3-lactamases (classes A and C) contain an active site serine and are thought to have evolved from the dd transpeptidases, but the B typey has a catalytic Zn2+. The latter, as well as a recently discovered type A enzyme,2 hydrolyze imipenem, currently one of the antibiotics of last resort used to treat infections by penicillin-resistant bacteria. Some (3-lactam antibiotics are also powerful inhibitors of (3-lactamases.U/aa/bb These antibiotics may also have uses in inhibition of serine proteasesCC/dd such as elastase. Some antibiotic-resistant staphylococci produce an extra penicillin-binding protein that protects them from beta lactams.ee Because of antibiotic resistance the isolation of antibiotics from mixed populations of microbes from soil, swamps, and lakes continues. Renewed efforts are being... 

ANTIBIOTICS 0-LACTAMASE INHIBITORS. The antibacterial effectiveness of penicillins, cephalosporins, and other 0-lactam antibiotics depends on selective acylation and consequently, inactivation, of transpeptidases involved in bacterial cell wall synthesis. This acylating ability is a result of the reactivity of the 0-lactam ring (1). Bacteria that are resistant to 0-lactam antibiotics often produce enzymes called 0-lactamases that inactivate the antibiotics by catalyzing the hydrolytic opening of the 0-lactam... 

The penicillin antibiotics inhibit transpeptidase enzymes (penicillin-binding proteins (PBPs)) by acylation of the serinyl residue at their active site, which leads to cell wall lysis, since blocking PBPs circumvents proper murein membrane formation [3]. Several peptides and peptidomimetics containing the (3-lactam ring have been recently described as effective protease inhibitors and, consequently, as potential drugs for a wide range of diseases implicating proteases [5-8]. 

Figure 5 A penicillin-binding activity-based high-throughput SPA assay for the identification of potential inhibitors of the transpeptidase activity of PBPs. A particular PBP of interest is first labeled with biotin. The biotinylated PBP is then mixed with a 3H-labeled P-lactam in 96-well microtiter plates and the reaction mixtures are incubated at room temperature for a period of time. After incubation, the biotionylated PBP-3H-P-lactam complexes are captured by mixing with streptavidin coated SPA beads. The P-lactam-binding activity of the PBP is measured by counting the microtiter plates after mixing with the SPA beads.

A cephalosporin derivative 257 with structural features of the peptidoglycan has been conceived as an inhibitor specific for DD-transpeptidases <2003JA16322>. The compound 257 has been synthesyzed in 13 steps and has been tested with recombinant PBPlb and PBP5 of E. coli, a DD-transpetidase and a DD-carboxypeptidase, respectively. It has been found that compound 257 is a time-dependent and irreversible inhibitor of PBPlb and does not interact with PBP5, neither as an inhibitor (reversible or irreversible) nor as a substrate. 

It is known that p-lactamase catalyzes the rapid hydrolysis of the p-lactam ring of penicillins and cepharosporines. The hydrolytic activity of these enzymes eliminates the bacteriocidal action of many p-lactam antibiotics and makes the organism resistant to these molecules. For this reason, the p-lactamase inhibitors have long been regarded as promising targets from a medicinal viewpoint. A comparison between the kinetic characteristics of p-lactamase and penicillin-sensitive enzymes (carboxy-peptidase and transpeptidase) is of interest in this respect. p-Lactamases very efficiently hydrolyze p-lactam in contrast to penicillin-sensitive enzymes [high /e4 in Eq. (9)]. 

Why is penicillin such an effective inhibitor of the transpeptidase The highly strained, four-membered P-lactam ring of penicillin makes it especially reactive. On binding to the transpeptidase, the serine residue at the active site attacks the carbonyl carbon atom of the lactam ring to form the penicilloyl-serine derivative (Figure 8.31). Because the peptidase participates in its own inactivation, penicillin acts as a suicide inhibitor. 

Finally, it may be worthwhile to point out that one of the most successful chemotherapeutic drugs provided by nature may be considered as a non-classical active-site-directed irreversible inhibitor This drug is penicillin (70). As a non-classical antimetabolite of D-alanyl-D-alanine, it first forms a reversible complex with the enzyme peptidoglycan transpeptidase, and then by a ringopening reaction of its p-lactam moiety, it forms a covalently linked penicil-loyl-enzyme complex97. However, the reaction involves the acylation of a sulfhydryl group in the active site of the enzyme in this respect, 70 resembles the classical-type endoalkylating antimetabolites, azaserine and DON (8 and 9, see Section 2.2.). Some of the more recently discovered antibiotics and natural products from plants with antitumor activity (e.g., camptothecin) are... 

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