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Enzymatic studies of the prenatal chick, with special reference to the multibple forms of liver soluble NADP-linked isocitric dehydrogenase

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Title Enzymatic studies of the prenatal chick, with special reference to the multibple forms of liver soluble NADP-linked isocitric dehydrogenase
Names Cheung, Geoffrey Peter (creator)
Newburgh, R. W. (advisor)
Date Issued 1966-08-12 (iso8601)
Note Graduation date: 1967
Abstract Levels of activity of glucose-6-phosphate- , 6-phosphogluconate- , and isocitric- dehydrogenase were measured in the
soluble and particulate fractions of heart, brain and liver at
several ages throughout the development of the chick embryo.
Compared to isocitric dehydrogenase, levels of glucosesphosphate-
, and 6-phosphogluconate- dehydrogenase are generally
low. This suggests that the majority of the NADPH in the prenatal
chick arises from the NADP-isocitric dehydrogenase. The results,
however, say nothing of the possibility of different pools of NADPH
arising from the different enzymes. A NADPH-NAD transhydrogenase
activity was found in the digitonin-treated mitochondrial
fractions of the same organs, and its levels were also determined.
They were found to be very low, making it unlikely that the majority
of the NADPH generated by the isocitric dehydrogenase is shunted into the electron transport chain of the mitochondria.
Multiple forms of the NADP-linked isocitric dehydrogenase
were shown to be present in the soluble fractions from liver of 18-
day old embryos, using differential rates of heat denaturation.
Partial purification of the soluble liver enzyme yielded two forms,
one stable and the other labile to heat. It was found that these two
enzymes have different sedimentation properties, pH optima and
reaction rates with NADP. The dimeric, heat stable form may be
dissociated into the monomeric, heat labile, form by a lowering
of the ionic strength. This dissociation is readily reversible, and
the newly formed dimer appears to be similar with the native
dimer.
On the basis of experiments presented and comparison with
the behavior of similar enzymes in other organisms, a mode of
interaction of the multiple forms is postulated. Soluble NADP-linked
isocitric dehydrogenase of liver may play a role in the
regulation of cellular metabolism during development.
Genre Thesis/Dissertation
Topic Embryology -- Birds
Identifier http://hdl.handle.net/1957/47256

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