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Peptide Based Artificial Receptors for Carbohydrate Anthrose Detection.

Anthrax is a fatal infection that occurs when Gram-positive Bacillus anthracis spores enter the mammalian host through abrasion in skin, inhalation or ingestion. B. anthracis spores are very resistant and can remain dormant in soil for decades. Therefore, an effective detection system for B. anthracis is urgently needed. Recently, it was found that one of the components of the B. anthracis exosporuim is a collagen like protein whose carbohydrate portion is composed of the tetrasaccharide with the highly specific monosaccharide upstream terminal, named anthrose. Since anthrose was not found on other bacterial spores, including those closely related to B. anthracis, this monosaccharide is an attractive target for the development of new B. anthracis detection and identification methods. Peptide cyclization represents particularly interesting approach for the design of artificial receptors for anthrose, because cyclic peptides provide the possibility of having a spherical lipophilic binding site of appropriate size and shape for a particular carbohydrate substrate. The presence of hydrogen donor/acceptor groups within a three-dimensional structure permits carbohydrate substrates to be encapsulated, thereby allowing their binding in water. In order to determine whether the cyclic peptide receptor can selectively detect the anthrose, we have successfully prepared cyclic peptide combinatorial library (total 6859 peptides) by the process of divide, couple and recombine (“tea-bag” technology) using standard Fmoc solid-phase peptide synthesis. Prepared combinatorial library is screened for anthrose binding in fluorescence-based assay, and individual cyclic peptides with enhanced affinity toward anthrose are identified by the positional scanning deconvolution process.

anthrax; cyclic peptide combinatorial library; receptors; solid-phase peptide synthesis; fluorescence-based assay

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