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Biomedical subjects

V Kasche

Publications and source records attributed to V Kasche.

At least 55 records · Page 3Linked to original sources
PubMed1981

A method for the isolation of homogeneous fluorescein-labeled proteins for studies on the variation in biological and fluorescence properties of the different conjugates.

Different homogeneous fluorescein-labeled conjugates of alpha-chymotrypsin and the pancreas trypsin inhibitor have been analyzed and isolated by isoelectric focusing with and without biospecific sample application. With this procedure the biological activity of the different protein conjugates can be determined from the focusing pattern. The fluorescence quantum yield was found to be decreased with the number of labels per protein. The biological activity, however, was increased in some monolabeled conjugates compared with the activity of the native protein. The quantum yield was found to be markedly (greater than 10%) changed when the monolabeled conjugates formed specific complexes with proteins.

Animals↗
PubMed1977

Rapid zymogen activation and isolation of serine proteases from an individual mouse pancreas by affinity chromatography: genetical heterogeneity of chymotrypsins of Mus musculus.

A rapid method to prepare homogeneous fractions of the various chymotrypsins and trypsins from a single mouse pancreas (130-150 mg wet weight) is described. The method was applied to investigate intra-species variation on a molecular level using chymotrypsins as biochemical indicators. The conditions for optimal extraction of the zymogens in the homogenized pancreas have been studied. DNA had to be removed from the homogenate to obtain maximum chymotrypsin yields (approximately 1% of the wet weight of the pancreas). The activation was initiated by immobilized bovine trypsin that was removed by filtration. Then chymotrypsinogens in the homogenate were activated by mouse trypsin. After completed activation homogeneous chymotrypsins (one anionic and one cationic form) could be isolated in an one step analytical affinity chromatographic separation, using soybean trypsin inhibitor bound in Sepharose as a protease specific adsorbent. The end products were characterized by isoelectric focussing, amino acid composition, enzymatic parameters, molar extinction coefficient, and the number of polypeptide chains. Hereby, the existence of two chymotrypsinogen loci in the mouse genome could be demonstrated. Differences in structure and function between the corresponding enzymes from the two strains were found. This allelomorphism was verified in the crossing of the off-spring.

Alleles↗
PubMed1976

Resolution in affinity chromatography. The effect of the heterogeneity of immobilized soybean trypsin inhibitor on the separation of pancreatic proteases.

By affinity chromatography, trypsins and chymotrypsins from mouse pancreas homogenates have been separated using soybean trypsin inhibitor immobilized on Sepharose. The effects of the functional heterogeneity of the adsorbent have been investigated in terms of the resolution obtained. Heterogeneity of the adsorbent have been investigated in terms of the resolution obtained. Heterogeneity has been found to originate from the following sources: heterogeneity of the ligand before immobilization; alteration of the ligand by immobilization; and modification of the ligand after immobilization by molecules to be fractionated. Only when the heterogeneity of the adsorbent was minimized could the resolution of closely related enzyme species be achieved. The elution conditions for different enzymes depended on the amount of enzyme applied, as no complete homogeneity could be obtained. In addition, it was found that the adsorbent was partly degraded by the pancreas extract, reducing its fractionating capacity.

Adsorption↗