Sequential analysis in subnanomolar amounts of peptides. Determination of the structure of a habituation-induced brain peptide (ameletin)

✍ Scribed by S.R. Burzynski

• Publisher: Elsevier Science
• Year: 1976
• Tongue: English
• Weight: 946 KB
• Volume: 70
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(76)90457-7

✦ Synopsis

Autoradiography of 3H-labeled dansyl amino acids was applied to the sequential analysis of peptides available only in subnanomolar quantities. The method employs time-course analysis of the digestion of the peptide by aminopeptidase M, carboxypeptidase B, and dipeptidylaminopeptidase I. The cleaved amino acids or dipeptides are identified by tic of the 3H-labeled compounds. The procedure was used for the sequential analysis of a learning-induced peptide extracted from trained rat brain, called ameletin. The sequence, confirmed by synthesis, was: p-Glu-Ala-Gly-Tyr-Ser-Lys.

MATERIALS AND APPARATUS

Standard amino acids were obtained from Calbiochem, standard dipeptides from Cyclo Chemical and Gallard-Schlesinger, and the synthetic hexapeptide H-Leu-Trp-Met-Arg-Phe-Ala-OH (peptide MS) from Research Plus Laboratories. Enzymes were purchased from the following sources: aminopeptidase M (AM), 5000 munit/mg, from Henly & Co. (manufactured by Rrhm GMBH Chemische Fabrik); carboxypep-359