Microsequence analysis of peptides and proteins: III. Artifacts and the effects of impurities on analysis

✍ Scribed by John E. Shively; David Hawke; Barry N. Jones

• Publisher: Elsevier Science
• Year: 1982
• Tongue: English
• Weight: 830 KB
• Volume: 120
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(82)90352-9

✦ Synopsis

Levels of contaminants in the parts-per-billion range can adversely affect amino acid microsequence analysis (low-nanomole to subnanomole range) in two ways; (a) contaminants in sobents used in the purification of proteins and peptides can derivatize reactive amino acids to form unusual products or react with free cr-NH2 groups to effectively prevent sequence analysis, and (b) contaminants in the reagents and solvents used in Edman chemistry can give spurious peaks on HPLC analysis of amino acid phenythiohydantoin derivatives or react with the phenylthiocarbamylpeptidyl derivatives to give lower initial and repetitive yields of the subsequent phenylthiohydantoin derivatives. Practical examples of these problems and their solutions are described. With proper care in the preparation of solvents and reagents for sample purification and Edman chemistry, microsequence analysis in the low-nanomole to subnanomole range can be made routine.