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G P Royer

Publications and source records attributed to G P Royer.

At least 19 recordsLinked to original sources
PubMed1980

Use of immobilized carboxypeptidase Y (I-CPY) as a catalyst for deblocking in peptide synthesis.

CPY is a metal-free carboxypeptidase from yeast with broad specificity [1]. In addition to exopeptidase activity at acid pH, the enzyme is an effective esterase at alkaline pH. N-alpha-acetyl-L-tyrosine ethyl ester is hydrolyzed faster by CPY than by chymotrypsin. These observations suggested that the immobilized form of the enzyme would be of value in removing ester groups from the C-terminal ends of peptides. In this report we describe sequential synthesis using I-CPY and alpha-COOH deblocking of peptides made by conventional methods.

Carboxypeptidases↗
PubMed1977

Substrate-induced conformational changes in lactate dehydrogenase. Proteolysis of the immobilized enzyme in the presence of specific substrates.

We report here a new approach to the study of the conformation of enzymes in the presence of specific substrates. Rabbit muscle lactate dehydrogenase was attached to CL-Sepharose via a cleavable spacer arm (-NH-(CH2)6NHCO(CH2)2SS(CH2)2CO-). The bound lactate dehydrogenase was digested with subtilisin BPN' in the presence of substrates of lactate dehydrogenase. The use of a flow system permits the maintenance of saturating levels of substrates. Proteolysis was followed by loss of activity of the enzyme column. The time course of proteolysis in the presence of either NADH, NAD+, or pyruvate alone did not differ from the control. However, when NADH and pyruvate were present simultaneously, the enzyme became more susceptible to proteolysis. The initial rate of proteolysis was increased by 40%. The abortive ternary complex (lactate dehydrogenase - NAD+ - pyruvate) also showed an increase in susceptibility to proteolysis. These findings clearly show that the productive ternary complex (lactate dehydrogenase - NADH - pyruvate) is conformationally different from the apoenzyme and binary complexes under optimal catalytic conditions.

Enzymes, Immobilized↗
PubMed1977

The effect of calcium ion on the urea denaturation of immobilized bovine trypsin.

The effect of calcium ion on the urea denaturation of trypsin has been investigated. By using trypsin immobilized on glass beads, all possibilities of autolysis occurring during the denaturation process are eliminated. It was found that in 8 M urea calcium ion markedly decreases the denaturation rate of the immobilized trypsin. Conversely, the presence of calcium ion markedly accelerates the rate of renaturation of denatured immobilized trypsin. Calcium may exert its stabilizing effect on the tertiary structure of the protein by coordination to the side chains of Asp 194, Ser 190 and the carbonyl group of Ser 139 (using the chymotryptic numbering system).

Animals↗