Instrumentation and applications of an automated C-terminal fragment peptide fractionator for C-terminal sequence analysis of proteins.

A novel automated C-terminal fragment peptide fractionator has been constructed. Digests with lysyl endopeptidase were covalently immobilized on p-phenylene diisothiocyanate polymer beads. Only the C-terminal fragment, which contains no lysyl residue, was liberated by cleavage at the first peptide bond of the immobilized fragment peptides with trifluoroacetic acid, and it was automatically collected. The whole procedure was automatically and precisely performed under microprocessor control in a nitrogen atmosphere. The resulting fragment was sequenced without further purification. Sequences of both N- and C-terminal regions can be routinely obtained for ca. 100 pmol samples by the use of a conventional automated Edman-type protein sequencer.