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Biomedical subjects

C L Wright

Publications and source records attributed to C L Wright.

13 recordsLinked to original sources

Purification of the hexokinases by affinity chromatography on sepharose-N-aminoacylglucosamine derivates. Design of affinity matrices from free solution kinetics.

The purification is described of rat hepatic hexokinase type III and kidney hexokinase type I on a large scale by using a combination of conventional and affinity techniques similar to those previously used for the purification of rat hepatic glucokinase [Holroyde, Allen, Storer, Warsy, Chesher, Trayer, Cornish-Bowden & Walker (1976) Biochem. J. 153, 363-373] and muscle hexokinase type II [Holroyde & Trayer (1976) FEBS Lett. 62, 215-219]. The key to each purification was the use of a Sepharose-N-aminoacylglucosamine affinity matrix in which a high degree of specificity for a particular hexokinase isoenzyme could be introduced by either varying the length of the aminoacyl spacer and/or varying the ligand concentration coupled to the gel. This was predicted from a study of the free solution kinetic properties of the various N-aminoacylglucosamine derivatives used (N-aminopropionyl, N-aminobutyryl, N-aminohexanoyl and N-aminooctanoyl), synthesized as described by Holroyde, Chesher, Trayer & Walker [(1976) Biochem. J. 153, 351-361]. All derivatives were competitive inhibitors, with respect to glucose, of the hexokinase reaction, and there was a direct correlation between the Ki for a particular derivative and its ability to act as an affinity matrix when immobilized to CNBr-activated Sepharose 4B. Muscle hexokinase type II could be chromatographed on the Sepharose conjugates of all four N-aminoacylglucosamine derivatives, although the N-aminohexanoylglucosamine derivative proved best. This same derivative was readily able to bind hepatic glucokinase and hexokinase type III, but Sepharose-N-amino-octanoyl-glucosamine was better for these enzymes and was the only derivative capable of binding kidney hexokinase type I efficiently. Separate studies with yeast hexokinase showed that again only the Sepharose-N-amino-octanoylglucosamine was capable of acting as an efficient affinity matrix for this enzyme. Implications of these studies in our understanding of affinity-chromatography operation are discussed.

Animals

Campylobacter fetus (Vibrio fetus) infection in dairy herds in South-West Scotland.

Infertility and abortion caused by Campylobacter fetus is described in a series of 12 herds in South West Scotland during the period May 1974 to December 1976. Herd studies show that 44 bulls were involved and that there were 34 bull and 18 cow movements for breeding purposes. The need to include C fetus infection in the differential diagnosis of herd infertility is emphasised.

Abortion, Veterinary

The pepsinogens of cat gastric mucosa and the pepsins derived from them.

Fundic mucosal extracts of the cat were examined by chromatography using DEAE-cellulose at pH 5.3. The peaks of proteolytic activity located by this method were shown to be heterogeneous on electrophoresis. By electrophoresis of individual homogenous chromatographic fractions 9 separate zones of proteolytic activity were detected. One zone did not migrate from the origin and is possibly a non-pepsinogen gastric protease. Simultaneous electrophoresis of pepsinogens and pepsins indicated that each pepsinogen gave rise to one pepsin. One zymogen gives rise to a pepsin active with the synthetic substrate acetyl-L-phenylalanyl-L-diiodotyrosine (APDT).

Animals

Variation in the proportions of individual pepsins secreted by the cat in response to vagal stimulation and hypoglycaemia.

1. Gastric juice was collected at regular intervals during electrical stimulation of the vagus in anaesthetized cats and during insulin hypoglycaemia in both anaesthetized and conscious cats. The total amounts of acid and pepsin secreted were similar in the three groups. 2. Pepsins were examined by agar-gel electrophoresis. Resting juice contained two pepsins, and up to nine pepsins could be detected after stimulation. Three patterns of pepsin secretion were found. 3. The most noticeable feature was the variation in the proportion of total pepsin attributable to the pepsin which migrated most rapidly during electrophoresis (pepsin 1). In response to insulin hypoglycaemia, anaesthetized cats secreted only a small proportion of total pepsin 1 and conscious cats secreted a large proportion as pepsin 1. During direct electrical stimulation of the vagus, the proportion of pepsin 1 rose. 4. The possibility of a dependence of pepsin 1 secretion on vagal stimulation is discussed and the relevance of this to peptic ulcer and to vagotomy is considered.

Animals