Summary
Methods are given in detail to assay branched chain keto acid oxidases in native leucocytes and fibroblasts. In peripheral blood these enzymes are located preferentially in lymphocytes.
The intraindividual variation of enzyme activities in leucocytes is reduced by correcting for the number of lymphocytes. In contrast, interindividual variation remains unchanged. Consequently, an overlap between enzyme activities of control persons and heterozygotes for classic maple syrup urine disease still exists. For explanation multiple alleles and influence of genetic background on enzyme activities are invoked.
Arguments are given for the simultaneous defect of the three branched chain keto acid oxidases in classic maple syrup urine disease.
Furthermore some new observations on the intermittent type of maple syrup urine disease are given.
Tests for heterozygosity in fibroblasts are complicated because of environmental influences in cultures which are not fully understood at present. However, the enzymatic defect is clearly demonstrated in fibroblasts of patients with the classic type and the intermittent type of maple syrup urine disease.
Zusammenfassung
Die Methoden zur Testung der Oxidasen für die verzweigtkettigen α-Ketosäuren in Leukocyten und Fibroblasten werden beschrieben. Im peripheren Blut sind diese Enzyme bevorzugt in den Lymphocyten lokalisiert.
In den Leukocyten wird die intraindividuelle Variation der Enzymaktivitäten durch Berücksichtigung des Differentialblutbildes verringert. Die interindividuelle Variation bleibt dagegen unverändert. — Für die Enzymaktivitäten von Normalpersonen und Eltern von Patienten mit klassischer Ahornsirupkrankheit bleibt damit ein Überlappungsbereich bestehen. Als mögliche Erklärung werden multiple Allelie und multifaktorielle Determinierung von Enzymaktivitäten diskutiert.
Bisher gewonnene Ergebnisse lassen vermuten, daß bei der klassischen Form der Ahornsirupkrankheit alle drei Oxidasen für die verzweigtkettigen α-Ketosäuren defekt sind. Neuere Untersuchungen über die intermittierende Form der Ahornsirupkrankheit werden mitgeteilt.
Die Erkennung von Heterozygoten in Testen mit Fibroblasten ist erschwert, da die Abhängigkeit der Aktivität der α-Ketosäure-Oxidasen von den Kulturbedingungen noch nicht genügend geklärt ist. Es ist dagegen möglich, Patienten mit der klassischen und der intermittierenden Form der Ahornsirupkrankheit durch enzymatische Teste an Fibroblasten zu erkennen.
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Dedicated to Prof. Dr. K. H. Schäfer at the occasion of his 60th birthday.
This work was supported in part by Deutsche Forschungsgemeinschaft and Stiftung Volkswagenwerk.
U. L. is recipient of a training grant from Stiftung Volkswagenwerk.
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Langenbeck, U., Rüdiger, H.W., Schulze-Schencking, M. et al. Evaluation of a heterozygote test for maple syrup urine disease in leucocytes and cultured fibroblasts. Hum Genet 11, 304–315 (1971). https://doi.org/10.1007/BF00278658
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DOI: https://doi.org/10.1007/BF00278658