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.
| Identifer | oai:union.ndltd.org:unt.edu/info:ark/67531/metadc503944 |
| Date | 12 1900 |
| Creators | Kung-Chao, Diana T.-Y. |
| Contributors | Tai, Hsin-Hsiung (Daniel), Gracy, Robert W., Yorio, Thomas |
| Publisher | North Texas State University |
| Source Sets | University of North Texas |
| Language | English |
| Detected Language | English |
| Type | Thesis or Dissertation |
| Format | iii, 86 leaves: ill., Text |
| Rights | Public, Kung-Chao, Diana T.-Y., Copyright, Copyright is held by the author, unless otherwise noted. All rights reserved. |
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