Summary. This investigation describes the quantitative and qualitative distribution
of lactate dehydrogenase (LDH) isoenzymes in subcellular particulate
fractions of tissues isolated by differential centrifugation. All fractions, especially
those containing microsomes and nuclei, had measurable LDH activity. The
supernatant remaining after centrifugation of the microsomal fraction contained
40—50% of the total LDH activity and had an isoenzyme pattern similar to that
found in the corresponding tissue homogenate. The cathodically migrating LDH
isoenzyme, LDH(5) (M(4)), was found mainly in nuclear fractions and the anodically migrating
isoenzyme, LDHX (H(4)), in purified mitochondrial fractions. By mixing in
vitro equal amounts of nuclear (LDH(5)) and mitochondrial (LDH(4)) fractions, intermediate
isoenzyme patterns were obtained containing LDH(2), LDH(3), and LDH(4). If
the nuclear and mitochondrial fractions were mixed in a ratio of 10/1 respectively,
then the isoenzyme pattern was similar to that seen in malignant tissue in general.
The results suggest that changes in the structural elements of the cell, i.e. nuclei
and mitochondria, involves changes in the LDH isoenzyme pattern. According to
this hypothesis the LDH pattern of malignant tissue is correlated with well-recognized
morphologic changes in malignant cells.
(1) A method for measurering the total activity of urinary lactate dehydrogenase (urinary LDH) is presented. The optima of pH, substrate and Coenzyme for this enzyme were determined by measuring the decrease in optical density at 366 nm of NADH formed by reduction of pyruvate to lactate. By application of this method no urinary inhibitor of LDH was detected.
(2) Urinary LDH activities were investigated in patients suffering from some severe renal diseases, including unilateral renal ischaemia. Simultaneously the urinary LDH-isozyme pattern was determined by means of zone electrophoresis in agar gel.
(3) The value of this supplement to the measurement of the total enzyme activity is pointed out. Thus unilateral renal ischaemia may be associated with the relative predominance of LDH-isozyme five in the urine from the affected kidney.
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