scholarly journals Studies on heterogeneity of human gastric zymogens

1970 ◽  
Vol 116 (3) ◽  
pp. 397-404 ◽  
Author(s):  
Michael D. Turner ◽  
Jagdish C. Mangla ◽  
I. Michael Samloff ◽  
Leon L. Miller ◽  
Harry L. Segal
Keyword(s):  
Gastric Mucosa ◽  
Gel Electrophoresis ◽  
Deae Cellulose ◽  
Human Gastric Mucosa ◽  
Active Components ◽  
Agar Gel ◽  
The Past ◽  
Agar Gel Electrophoresis ◽  
Gradient Chromatography

The peptic zymogens of human gastric mucosa have been studied in the past by three different techniques: phosphate-gradient chromatography on a column of DEAE-cellulose, chloride-gradient chromatography on DEAE-cellulose and agar-gel electrophoresis. These techniques have given different results, each suggesting the presence of a different number of pepsin zymogens in human gastric mucosa. In the present experiments gastric mucosal homogenates were subjected to analysis by all three techniques. The two major zymogen peaks obtained from phosphate-gradient chromatography were found to be composed of the three fractions found on chloride-gradient chromatography; these fractions in turn were shown to be heterogeneous on agar-gel electrophoresis. The latter technique demonstrated the presence of seven separable enzymically active components in human gastric mucosal homogenates. The relationships of the components separated by the three techniques are discussed.

The Proteinases of Human Gastric Adenocarcinomata: Their Identification, Separation and Sites of Action on the B-Chain of Oxidized Insulin

1972 ◽  
Vol 42 (1) ◽  
pp. 79-90 ◽  
Author(s):  
D. J. Etherington ◽  
W. H. Taylor
Keyword(s):  
Column Chromatography ◽  
Gel Electrophoresis ◽  
Deae Cellulose ◽  
Intermediate Form ◽  
Agar Gel ◽  
Equivalent Position ◽  
Normal Tissues ◽  
Highly Active ◽  
Agar Gel Electrophoresis ◽  
Sites Of Action

1. Because of the possibility that the proteinases of gastric adenocarcinomata may differ from those of healthy gastric mucosa, an investigation of these enzymes by means of pH-activity studies, agar-gel electrophoresis, column chromatography and the mode of action on the B-chain of oxidized insulin has been undertaken. 2. Agar-gel electrophoresis of extracts at pH 5 revealed a single zone of proteinase activity situated at the equivalent position to the zone 7 of normal gastric mucosal extracts and not activated at pH 2. The typical pepsins of normal mucosal extracts were not present. 3. Agar-gel electrophoresis at pH 8·2 resolved this single zone into three zones. One was located slightly cathodally (proteinase 1) and the other two anodally (a slower proteinase 2A and a faster 2B). 4. Proteinases 2A and 2B were separated by column chromatography and shown to have identical asymmetrical broad pH-activity curves with the maximum at pH 3·3–3·4. 5. Proteinase 1 had a symmetrical narrow pH-activity curve with the maximum at pH 3·7–3·9. Proteinase 1 was purified and resolved into two highly active major components, 1A and 1B, by column chromatography, first on DEAE-cellulose and then on CM-cellulose. 6. The sites of cleavage of the B-chain of oxidized insulin were determined for proteinases 1A and 1B. The same bonds were split by each, with one exception, but several were split at differing rates indicating that the two enzymes had related, but different, modes of action. 7. The tumour proteinases 1 resemble the cathepsins D in certain respects and the pepsins in others. They may represent an enzyme structure of an intermediate form. There is insufficient evidence to indicate whether they are elaborated by partially differentiated cells or whether they are derived from cells which have become dedifferentiated. No enzymes exactly like them have yet been found in normal tissues.

Isolation and some properties of 11- and 9-Gla prothrombins from normal plasma

1991 ◽  
Vol 69 (5-6) ◽  
pp. 404-408
Author(s):  
Om P. Malhotra
Keyword(s):  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Deae Cellulose ◽  
Agar Gel ◽  
Bovine Plasma ◽  
Carboxyglutamic Acid ◽  
K Deficiency ◽  
Agar Gel Electrophoresis ◽  
Relationship Of

The relationship of prothrombin structure to function with respect to γ-carboxyglutamic acid (Gla) residues can be effectively evaluated by characterizing the behavior of prothrombin isomers differing in Gla content. In addition to the isolation of a whole spectrum of Gla-deficient, 0- to 9-Gla isomers from dicoumarol-treated plasma, prothrombin isomers containing 11 (10.90) and 9 (8.85) Gla residues have now been isolated from normal bovine plasma. The isomers were isolated by barium citrate adsorption, elution, and finally by heparin-agarose, DEAE-cellulose, and immuno-affinity chromatographies. Each of the purified isomers showed a single component by agar gel and sodium dodecyl sulfate – polyacrylamide gel electrophoresis. By agar gel electrophoresis, the 11-Gla prothrombin isomer moved the fastest, followed by the 10-, and lastly the 9-Gla isomer, independent of Ca2+. The corresponding 9-, 10-, and 11-Gla prothrombin fragments 1 exhibited similar migration tendencies. By gel electrofocusing, 11- and 9-Gla fragments 1, respectively, focused anodal and cathodal to 10-Gla fragment 1. The Ca2+-induced decrease in the intrinsic fluorescence in 11-, 10-, and 9-Gla fragments 1 was 48, 40, and 45%, respectively. This metal-induced structural change did not correlate with the functional, thrombin-generating property of the isomers, as the 9-Gla variant exhibited 75%, and the 11-Gla 110–115%, of normal coagulant activity.Key words: prothrombin, blood clotting, dicoumarol, Warfarin, γ-carboxyglutamic acid, vitamin K deficiency.

scholarly journals The preparation and purification of individual human pepsins by using diethylaminoethyl-cellulose

1978 ◽  
Vol 169 (3) ◽  
pp. 607-615 ◽  
Author(s):  
N B Roberts ◽  
W H Taylor
Keyword(s):  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Deae Cellulose ◽  
Preliminary Evidence ◽  
Peptic Activity ◽  
Agar Gel ◽  
Diethylaminoethyl Cellulose ◽  
Agar Gel Electrophoresis ◽  
Individual Human

A procedure was devised for isolating human pepsins 1, 2, 3 and 5 from gastric juice by repetitive column chromatography on DEAE-cellulose. The combined yields in four different experiments varied from 14% to 90% of the total peptic activity of the starting material. The isolated individual pepsins were shown to behave as single homogeneous proteins on agar-gel electrophoresis at pH 5.0 and on sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. There is preliminary evidence, requiring further study, of two other pepsins, one migrating between pepsins 1 and 2 and the other a pepsin-3 component associating closely with pepsin 5 on chromatography.

POLYETHYLENE GLYCOL ON AGAR-GEL ELECTROPHORESIS

The Lancet ◽  
1974 ◽  
Vol 304 (7892) ◽  
pp. 1321-1322
Author(s):  
W.H. Taylor ◽  
D.J. Etherington
Keyword(s):  
Polyethylene Glycol ◽  
Gel Electrophoresis ◽  
Agar Gel ◽  
Agar Gel Electrophoresis
Sign in / Sign up

Export Citation Format

Share Document