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http://hdl.handle.net/11375/22772
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DC Field | Value | Language |
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dc.contributor.advisor | Harrison, Paul | - |
dc.contributor.author | Bell , Karen Lesley | - |
dc.date.accessioned | 2018-04-27T18:26:06Z | - |
dc.date.available | 2018-04-27T18:26:06Z | - |
dc.date.issued | 1993-08 | - |
dc.identifier.uri | http://hdl.handle.net/11375/22772 | - |
dc.description.abstract | Hydroxymethylglutaryl-CoA synthase (HMG-CoA synthase) catalyzes the formation of HMG-CoA from acetyl-CoA and acetoacetyl-CoA. F-244 (1), a naturally occurring J3-lactone isolated from Fusarium sp. ATCC 20788 and other species, is known to be a potent and specific inhibitor ofHMG-CoA synthase isolated from rat liver. 1Ji,, HO,.,._.. .. , '• .. ,....13_.,., ;=( 0 14 0 0 (1) (2) This thesis describes the 48 fold purification of HMG-CoA synthase from bakers yeast in a three step procedure involving ethanol fractionation followed by ammonium sulfate precipitation and then hydroxylapatite chromatography. This procedure was found to be reproducible and yields a preparation of specific activity 0.14 units (j...tmolfmin)/mg in an overall yield of 8%. In our study, F-244 was found to be a potent irreversible inhibitor of HMGCoA synthase isolated from bakers yeast, with an IC50 value of 0.009 j...tM. This value is almost identical to the inhibitory activity of F-244 on rat liver HMG-CoA synthase that has been reported in the literature. Tritium labeled F-244 was prepared, for the first time, by feeding methyl-[3H]methionine to cultures of Fusarium sp. The [15, 16, 17, 18-3H] F-244 isolated had a specific activity of 1.3 x 106 DPM/mg. This tritiated F-244 was then used as an affinity.label for HMG-CoA synthase. Attempts to isolate the enzyme-inhibitor complex were unsuccessful due to the low level of radioactivity associated with the tritiated F-244. HMG-CoA synthase was also shown to be inhibited in a time-dependent irreversible manner by {±)-(3-butyrolactone (2). The rate of inactivation (~) was found to be 0.4697 s-1 and the inhibition constant (K1) was found to be 9 mM. The inactivation was found to be irreversible over several hours. | en_US |
dc.language.iso | en_US | en_US |
dc.subject | synthase, purification, ethanol fractionation, irreversible | en_US |
dc.title | Purification and Inhibition of Hydroxymethylglutaryl Coenzyme A Synthase | en_US |
dc.type | Thesis | en_US |
dc.contributor.department | Chemistry | en_US |
dc.description.degreetype | Thesis | en_US |
dc.description.degree | Master of Science (MS) | en_US |
Appears in Collections: | Digitized Open Access Dissertations and Theses |
Files in This Item:
File | Description | Size | Format | |
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Bell_Karen_L_1993Aug_Masters.pdf | 2.94 MB | Adobe PDF | View/Open |
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