NAD+-Dependent 15-Hydroxyprostaglandin Dehydrogenase from Swine Kidney: Characterization and Kinetic Mechanism

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Cytoplasmic 15-hydroxyprostaglandin dehydrogenase from swine kidney was purified to specific activity of 1.2 U per mg protein, by chromatographic techniques. Native molecular weight of enzyme was estimated at 45,000. Enzyme was inhibited by sulfhydryls, diuretics, and various fatty acids. Substrate studies indicated NAD+ specificity and ability to catabolize prostaglandins, except prostaglandin B and thromboxane B. Initial velocity studies gave intersecting plots conforming to a sequential mechanism. 15-keto-prostaglandin exhibited linear noncompetitive production inhibition with respect to either prostaglandin or NAD+; NAD yielded linear competitive production inhibition with respect to NADH. Results, and those of dead-end inhibition and alternated substrate studies, are ... continued below

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iii, 86 leaves: ill.

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Kung-Chao, Diana T.-Y. December 1979.

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  • Kung-Chao, Diana T.-Y.

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Cytoplasmic 15-hydroxyprostaglandin dehydrogenase from swine kidney was purified to specific activity of 1.2 U per mg protein, by chromatographic techniques. Native molecular weight of enzyme was estimated at 45,000. Enzyme was inhibited by sulfhydryls, diuretics, and various fatty acids. Substrate studies indicated NAD+ specificity and ability to catabolize prostaglandins, except prostaglandin B and thromboxane B. Initial velocity studies gave intersecting plots conforming to a sequential mechanism. 15-keto-prostaglandin exhibited linear noncompetitive production inhibition with respect to either prostaglandin or NAD+; NAD yielded linear competitive production inhibition with respect to NADH. Results, and those of dead-end inhibition and alternated substrate studies, are consistent with an ordered Bi-Bi mechanism: NAD+ is added first, then prostaglandin; then 15-keto-rostaglandin is released, then NADH.

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iii, 86 leaves: ill.

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  • December 1979

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  • May 10, 2015, 6:16 a.m.

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  • June 13, 2016, 2:17 p.m.

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Kung-Chao, Diana T.-Y. NAD+-Dependent 15-Hydroxyprostaglandin Dehydrogenase from Swine Kidney: Characterization and Kinetic Mechanism, thesis, December 1979; Denton, Texas. (digital.library.unt.edu/ark:/67531/metadc503944/: accessed June 21, 2018), University of North Texas Libraries, Digital Library, digital.library.unt.edu; .