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Purification and some properties of 11-hydroxythromboxane B2 dehydrogenase from porcine kidney
| Content Provider | Scilit |
|---|---|
| Author | Westlund, Pär |
| Copyright Year | 1993 |
| Description | Journal: Journal of Biological Inorganic Chemistry A protein with NAD-dependent 11-hydroxythromboxane B2 dehydrogenase activity was purified to apparent homogeneity from porcine kidney using a relatively simple purification procedure, involving precipitation, anion-exchange chromatography (diethylaminoethyl-cellulose), affinity chromatography (5'-AMP-Sepharose) and gel-filtration chromatography (Protein Pak 125). The dehydrogenase was found to have a molecular mass of 50-55 kDa as determined by comparison with standards on SDS/PAGE. The molecular mass on gel-filtration chromatography was dependent on the ionic strength of the buffer. The apparent Km and Vmax values for thromboxane B2 were also dependent of the ionic strength with a Vmax of 214 nmol min-1 mg-1 using 250 mM Tris/HCl, pH 8.0, and a corresponding Km of 2.9 mM. The enzyme was NAD dependent and was clearly separated from the proteins with 15-hydroxyprostaglandin dehydrogenase activity also present in the kidney. Furthermore, it was found that 11-hydroxythromboxane B2 dehydrogenase did not utilize prostaglandin D2, prostaglandin E2, prostaglandin F2 alpha or cholic acid as substrate, and that the enzyme did not catalyse the reverse reaction, conversion of 11-dehydrothromboxane B2 to thromboxane B2. |
| Related Links | http://onlinelibrary.wiley.com/doi/10.1111/j.1432-1033.1993.tb17675.x/pdf |
| Ending Page | 409 |
| Page Count | 7 |
| Starting Page | 403 |
| ISSN | 09498257 |
| e-ISSN | 14321327 |
| DOI | 10.1111/j.1432-1033.1993.tb17675.x |
| Journal | Journal of Biological Inorganic Chemistry |
| Issue Number | 2 |
| Volume Number | 212 |
| Language | English |
| Publisher | Wiley-Blackwell |
| Publisher Date | 1993-03-01 |
| Access Restriction | Open |
| Subject Keyword | Journal: Journal of Biological Inorganic Chemistry Analytical Chemistry Hydroxythromboxane B2 Dehydrogenase Dehydrogenase From Porcine |
| Content Type | Text |
| Resource Type | Article |
| Subject | Biochemistry Inorganic Chemistry |
