EC Number   |
|---|
    2.3.1.9 | - |
| 2.3.1.9 | acetoacetyl-CoA thiolase and 3-hydroxy-3-methylglutaryl -CoA synthase HMGCS form a complex. HMGCS catalyzes the second reaction in the mevalonate pathway. The 380-kDa crystal structure reveals that both enzymes are held together by a third protein (DUF35) with so-far-unknown function. The active-site clefts of thiolase and HMGCS form a fused CoA-binding site, which allows for efficient coupling of the endergonic thiolase reaction with the exergonic HMGCS reaction. The thiolase/HMGCS complex alone is able to convert acetyl-CoA to HMG-CoA. The tripartite complex is found in almost all archaeal genomes and in some bacterial ones |
| 2.3.1.9 | hangig-drop vapor diffusion at 21°C, crystal structure at 2.0 A resolution |
| 2.3.1.9 | hanging drop vapor diffusion method at 4°C. Unliganded and liganded (with CoA and with K+) structures of the human mitochondrial recombinant tetrameric thiolase |
| 2.3.1.9 | hanging drop vapor diffusion method, using 100 mM phosphate-citrate pH 4.2, 10% (w/v) polyethylene glycol 3350, 200 mM sodium chloride |
| 2.3.1.9 | microbatch method, using either 20% (w/v) PEG 3350, 0.15 M calcium chloride dehydrate, or 45% (w/v) PEG 200, 0.1 M MES monohydrate pH 6.0, 0.07 M calcium chloride dehydrate or 0.2 M sodium acetate trihydrate, 0.1 M sodium cacodylate trihydrate pH 6.5, 30% (w/v) PEG 8000, 5% (v/v) n-octyl-beta-D-glucoside |
| 2.3.1.9 | native enzyme at 1.8 A resolution, in complex with acetyl-CoA, at 1.9 A resolution |
| 2.3.1.9 | only successful in the presence of CoA |
| 2.3.1.9 | purified recombinant enzyme in apoform and with bound CoA, hanging drop vapor diffusion method, mixing 0.001 ml of 40 mg/ml protein in 40 mM Tris-HCl, pH 8.0, and 5 mM 2-mercaptoethanol, with 0.001 ml of reservoir solution containing 1.0 M ammonium sulfate, 0.1 M HEPES, pH 7.25, and equilibration against 0.5 ml of reservoir solution, 20°C, 3 days, X-ray diffraction structure determination and analysis at 2.0-2.3 A resolution, molecular replacement method using the structure of Mycobacterium tuberculosis thiolase Mttt0182, PDB ID1ULQ, as a search model, structure modeling |
| 2.3.1.9 | purified recombinant enzyme in apoform or with bound CoA, hanging drop vapor diffusion method, mixing 0.0012 ml of 25 mg/ml protein in 40 mM Tris-HCl, pH 8.0, with 0.0012 ml of reservoir solution containing 17% PEG 8000, 0.1 M HEPES pH 7.0, and equilibration against 0.5 ml of reservoir solution, 20-22°C, 7 days, X-ray diffraction structure determination and analysis at 1.4-1.5 A resolution, molecular replacement method using the structure of Mycobacterium tuberculosis thiolase MtFadA5, PDB ID 4UBU as a search model, structure modeling |
