Antimicrobial proteins

OxazoHdines, formed by reaction of alkanolamines with aldehydes, ate useful as leather tanning agents and ate effective curing agents for proteins, phenoHc resins, moisture-cure urethanes, etc. They also find use as antimicrobial agents. 

In subsequent studies attempting to find a correlation of physicochemical properties and antimicrobial activity, other parameters have been employed, such as Hammett O values, electronic distribution calculated by molecular orbital methods, spectral characteristics, and hydrophobicity constants. No new insight on the role of physiochemical properties of the sulfonamides has resulted. Acid dissociation appears to play a predominant role, since it affects aqueous solubiUty, partition coefficient and transport across membranes, protein binding, tubular secretion, and reabsorption in the kidneys. An exhaustive discussion of these studies has been provided (10). 

Astringents are designed to dry the skin, denature skin proteins, and tighten or reduce the size of pore openings on the skin surface. These products can have antimicrobial effects and are frequendy buffered to lower the pH of skin. They are perfumed, hydro-alcohoHc solutions of weak acids, such as tannic acid or potassium alum, and various plant extracts, such as bitch leaf extract. The alcohol is not only a suitable solvent but also helps remove excess sebum and soil from the skin. After-shave lotions generally function as astringents. 

Antibiotics are produced by fermentation. The process may take a few days to obtain an extractable amount of product. Antibiotic production is done by the batch process. Oxygen transport is the major concern therefore sufficient polymeric sugar and protein with a trace amount of elemental growth factors are used to enhance production. An anti-biogram test is used to observe the amount of antimicrobial agent in the fermentation broth. A bioassay determines the activity unit of the bactericides. 

Oxazolidinones are a new class of synthetic antimicrobial agents, which have activity against many important pathogens, including methicillin-resistant Staphylococcus aureus and others. Oxazolidinones (e.g. linezolid or eperezolid) inhibit bacterial protein synthesis by inhibiting the formation of the 70S initiation complex by binding to the 50S ribosomal subunit close to the interface with the 3OS subunit. 

Fig. 4. Comparison of the two signal-reaction chains leading either to the UV light-induced formation of flavonoids or to the elicitor-induced formation of furanocoumarins and related compounds with antimicrobial activity. From Hahlbrock et al. (1985). PR -proteins are pathogenesis-related proteins.

Notwithstanding the aforementioned difficulty in detecting specific target proteins other than the types normally observed in the taxonomic fingerprints from whole bacteria MALDI spectra (i.e., ribosomal proteins), some other target proteins and protein-like materials have been studied directly from whole cells. For example, Lantibiotics, antimicrobial peptides secreted by Gram-positive bacteria have been detected directly from whole bacteria by MALDI-TOF MS.51 The lantibiotics nisin and lacticin 481 were detected from whole cells and crude supernatants. Surprisingly, better results were reported from whole cells than the supernatants. In this study the presence of variants... 

In another study, the carrier protein was replaced by an enzyme compatible solid-phase resin (PEGA), and enzyme-catalyzed cyclization was used to probe substrate specificity. This study demonstrated also that oxo-esters are tolerated as substrates for TE domains, and then-preparation in library format served as an excellent tool for substrate specificity studies, as well as for preparation of cyclized peptides. Figure 13.11 shows how the TycA TE showed selectivity for only residues 1 and 9 (colored in red), and changes at all other residues were tolerated [42]. Hydrogen bonding interactions are shown in green. Several compounds made from this series were shown to demonstrate improved therapeutic indices (with respect to hemolysis) while retaining antimicrobial activity. 

Fig. 6 Illustration of representative cell envelopes from prokaryotes and eukaryotes that are typically encountered by antimicrobial peptides. The key components of the biomembranes and cell wall or giycocaiyx are shown, and the averaged protein content and typical dimensions are drawn to scale...

Unlike other Eukarya, animal cells lack cell walls, though they tend to be surrounded by a highly developed glycocalyx of up to 140 nm in thickness [108]. This diffuse layer of densely packed oligosaccharides has a heterogeneous composition and is connected to the membrane via lipids or integral proteins. The boundary of the cell usually extends beyond the mere lipid bilayer with its embedded proteins, and the extracellular structures provide initial sites of interaction or are themselves targets for MAPs such as antimicrobial peptides [115]. 

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