Untersuchungen zur histochemischen Darstellung der Sorbitdehydrogenase

Zusammenfassung Eine histochemische Nachweismethode für das zu den Polyoldehydrogenasen gehörende Sorbit-dehydrierende Enzym wird beschrieben. Insbesondere wird die Abgrenzung gegenüber der NAD-abhängigen Xylitdehydrogenase untersucht, da beide Fermente hinsichtlich ihres Wirkungsbereiches homospezifisch sind. Na-Aedta, das keinen Einfluß auf den histochemischen Nachweis der NAD-Xylitdehydrogenase besitzt, führt zu einer Verzögerung der histochemischen Darstellung der Sorbitdehydrogenase. Außerdem wird die Einwirkung von Natriumborat auf beide Fermentreaktionen untersucht, da diese Substanz biochemisch eine Hemmung der Sorbitdehydrogenase verursacht. Die mögliche Unterscheidung beider Enzyme auf Grund ihres Verhaltens wird erwogen. Abschließend wird die intrazelluläre Verteilung der beiden Polyoldehydrogenasen verglichen.

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Summary A topochemical method is described for the determination of the NAD-linked sorbitol-dehydrating enzyme. Furthermore it is discussed the relation of the sorbitoldehydrogenase to the NAD-linked xylitoldehydrogenase, as both enzymes are homospecific polyoldehydrogenases. Na₂-Edta has no influence on the determination of (NAD)-xylitoldehydrogenase, whereas a marked delay of the sorbitoldehydrogenase reaction is observed. Sodium borate as a biochemical inhibitor of the sorbitoldehydrogenase is also effective in the histochemical reaction and gives the base of a possible distinction between both enzymes. The intracellular distribution of the polyoldehydrogenases is compared.

     

The HLA-DRw8 lineage was generated by a deletion in the DR B region followed by first domain diversification

Sequences were generated for the first, second, and 3'UT regions of DRw8 beta-chain genes from two cell lines differing in their T cell determined allospecificities. Both have second domain sequences homologous to the B1 locus of the DRw52 family (DR3, DR5, and DRw6) and not the B3 locus. However, the 3'UT sequence is homologous to the 3'UT region of the B3 locus of the DRw52 family, and not the B1 locus. The first domain sequences are B1-like as opposed to B3-like and show polymorphism in the region encoding the putative alpha-helical region of the DR beta-chain. The easiest interpretation is that the DRw8 haplotypes constitute a sublineage within the DRw52 group and that this lineage has arisen by a small chromosomal deletion of the region between the B1 locus and the B3 locus. This deletion included the 3'UT region of the B1 locus, the B2 pseudogene, and the 5' end of the B3 locus including the exons encoding the first and second domains. After the deletion, two changes in the first domain arose by a mutational mechanism, possibly gene conversion. One of these changes parallels one seen in the DRw11 lineage.