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State-of-the-Art Peptide Synthesis: Comparative Characterization of a 16-mer Synthesized in 31 Different Laboratories

AddTime:2019-08-16

The Association of Biomolecular Core Facilities (ABRF) has 184 core facilities engaged in the synthesis and/or structural analysis of proteins and/or nucleic acids. custom peptide synthesis is a combination of several functions including synthesis, cleavage, purification, and characterization of the final product. Each of these functions poses specific problems that impact the quality of the final product. The mechanism for evaluating the interrelationship of these various functions was to ask that the member laboratories synthesize a 16 residue peptide of predetermined sequence by their preferred chemistry. Analysis of the purified product provides a monitor of purification techniques. Each of the 61 samples that was received were subjected to a battery of analytical techniques including amino acid analysis, HPLC, capillary electrophoresis, mass spectrometry, and biological assay as the peptide was designed to be a substrate for at least two different protein kinases. It was anticipated that these analyses would help to identify optimal methods for characterizing synthetic peptides as well as provide data regarding common side-products and reactions that may occur during custom peptide synthesis.

 These large, cylindrical complexes contain a central cavity that binds to unfolded polypeptides and sequesters them from the cellular environment. Substrate folding then occurs in this central cavity in an ATP-dependent manner. The eukaryotic chaperonin TCP-1 ring complex (TRiC, also called CCT) is indispensable for cell survival because the folding of an essential subset of cytosolic proteins requires TRiC, and this function cannot be substituted by other chaperones. This specificity indicates that TRiC has evolved structural and mechanistic features that distinguish it from other chaperones. Although knowledge of this unique complex is in its infancy, we review recent advances that open the way to understanding the secrets of its folding chamber.


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