PPT – PEPTIDE SYNTHESIS PowerPoint presentation | …
Presentation Summary : Peptides in chemistry and biology Phage display Solid phase peptide synthesis and applications of synthetic libraries Native chemical ligation in protein synthesis
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Presentation Summary : Daniel H. Rose, Jeremy J. Weyer, & Thao Yang Abstract In this project we employed the Solid Phase Peptide Synthesis Method to synthesize mucin peptides.
II.4.3.2 Inhibition of protein synthesis The initial observation that the trichothecene mycotoxins inhibited protein synthesis in mammalian cells was made by Ueno et al.
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As mentioned above, the generation and disappearance of Fmoc based chromophors allows the monitoring of the synthesis. Furthermore, samples may be taken to determine the load of Fmoc peptide. The completion of the deprotection reaction may be checked by cleaving samples and analyzing the obtained peptide.
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Choose an appropriate solvent, compatible with your experimental procedures and that does not react with or degrade the peptide. The solvent should be dry (if it is nonaqueous) and degassed. Since most peptide work is small-scale, it is most practical to buy small volumes of dry solvent, rather than attempting to dry it yourself. Determine if the peptide is acidic, basic or neutral (this depends on the ratio of different types of amino acid present in the sequence, and can be calculated using a range of free online tools, or the table in this guide). Proceed with solubilization using a small amount of peptide. Most acidic and basic peptides are more soluble at neutral pH. Peptides are acidic when the manufacturing process is complete, due to the presence of trifluoroacetate (TFA) as the counterion (a result of the cleavage or purification process). Check whether the peptide is supplied as a TFA salt before you start the solubilization process. Short non-hydrophobic peptides (25% non clustered, charged residues and
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But, whichever cleavage reagent is preferred, it has to be washed out very carefully after Fmoc removal, the last washing must be neutral. When synthesizing large peptides the duration of Fmoc cleavage should be gradually increased. For safe removal of the deblocking reagent the resin may have to be washed more often.
Protein Synthesis Translation - PowerPoint PPT Presentation
Also small scale manual SPPS as well as multiple peptide synthesis and the synthesis of pep- tide libraries can be performed very rapidly and conveniently with preformed active esters. Fmoc-AA-ONp and Fmoc-AA-OSu have found only restricted application in SPPS.