The Preparation and Primary Structure of S-Peptides from Different Pancreatic Ribonucleases
CORE Metadata, citation and similar papers at core.ac.uk Provided by Elsevier - Publisher Connector Volume 40, number 1 FEBS LETTERS March 1974 THE PREPARATION AND PRIMARY STRUCTURE OF S-PEPTIDES FROM DIFFERENT PANCREATIC RIBONUCLEASES G.W. WELLING, G. GROEN, D. GABEL+, W. GAASTRA, J.J. BEINTEMA Biochemisch Laboratorium, Rijksuniversiteit, Zernikelaan, Groningen, The Netherlands Received 14 December 1973 1. Introduction Miles-Seravac Ltd. (Maidenhead). All other ribonu- cleases used in this study (goat, giraffe, gnu, reindeer, In 1955, Richards [l] described the isolation of dromedary, kangaroo, lesser rorqual, pig, and horse) ‘an active intermediate produced during the digestion were isolated according to Wierenga et al. [7] and rat of ribonuclease by subtilisin’. The characterisation RNase, according to Beintema et al. [8]. Subtilopep- and separation of the non-covalently linked compo- tidase A (Subtilisin Carlsberg) was a gift from Novo nents was described 4 years later [2] . Ribonuclease Industri (Copenhagen). Sephadex G-50 (fine) was S* possesses full enzymatic activity and the same purchased from Pharmacia (Uppsala). All other rea- holds for the enzyme reconstituted from S-peptide gents were analytical grade products from Merck AG and S-protein. The involvement of S-peptide residues (Darmstadt). in the binding of S-peptide to S-protein and in the Amino acid analysis, high-voltage paper electro- enzymatic activity of the reconstituted RNase S’ has phoresis, dansylation, and dansyl-Edman degrada- been studied by using synthetic S-peptide analogs [3,4] tion were performed as described earlier [7, 93. the cleavage by subtilisin takes place in an external loop. Klee [5] and Gold [6] did not succeed in 2.1.
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