反応式
[CH3:1][C@@:2]([OH:58])([CH2:7][C:8]([S:10][CH2:11][CH2:12][NH:13][C:14]([CH2:16][CH2:17][NH:18][C:19]([C@H:21]([OH:57])[C:22]([CH2:25][O:26][P:27]([O:30][P:31]([O:34][CH2:35][C@H:36]1[O:40][C@@H:39]([N:41]2[C:45]3[N:46]=[CH:47][N:48]=[C:49]([NH2:50])[C:44]=3[N:43]=[CH:42]2)[C@H:38]([OH:51])[C@@H:37]1[O:52][P:53]([OH:56])([OH:55])=[O:54])([OH:33])=[O:32])([OH:29])=[O:28])([CH3:24])[CH3:23])=[O:20])=[O:15])=[O:9])[CH2:3][C:4]([OH:6])=[O:5].[CH:59]1[N:60]=[C:61]([NH2:106])[C:62]2[N:67]=[CH:66][N:65]([C@@H:68]3[O:72][C@H:71]([CH2:73][O:74][P:75]([O:78][P:79]([O:82][CH2:83][C@H:84]4[O:88][C@@H:87]([N:89]5[CH:94]=[C:93]([C:95]([NH2:97])=[O:96])[CH2:92][CH:91]=[CH:90]5)[C@H:86]([OH:98])[C@@H:85]4[OH:99])([OH:81])=[O:80])([OH:77])=[O:76])[C@@H:70]([OH:100])[C@H:69]3[O:101][P:102]([OH:105])([OH:104])=[O:103])[C:63]=2[N:64]=1>>[C:4]([O-:6])(=[O:5])[CH2:3][C@:2]([CH2:7][CH2:8][OH:9])([CH3:1])[OH:58].[CH:91]1[CH:90]=[N+:89]([C@H:87]2[O:88][C@@H:84]([CH2:83][O:82][P:79]([O:78][P:75]([O:74][CH2:73][C@H:71]3[O:72][C@@H:68]([N:65]4[C:63]5[N:64]=[CH:59][N:60]=[C:61]([NH2:106])[C:62]=5[N:67]=[CH:66]4)[C@H:69]([O:101][P:102]([OH:104])([OH:105])=[O:103])[C@@H:70]3[OH:100])([OH:77])=[O:76])([O-:81])=[O:80])[C@H:85]([OH:99])[C@@H:86]2[OH:98])[CH:94]=[C:93]([C:95]([NH2:97])=[O:96])[CH:92]=1.[CH3:24][C:22]([C@@H:21]([OH:57])[C:19]([NH:18][CH2:17][CH2:16][C:14]([NH:13][CH2:12][CH2:11][SH:10])=[O:15])=[O:20])([CH2:25][O:26][P:27]([O:30][P:31]([O:34][CH2:35][C@H:36]1[O:40][C@@H:39]([N:41]2[C:45]3[N:46]=[CH:47][N:48]=[C:49]([NH2:50])[C:44]=3[N:43]=[CH:42]2)[C@H:38]([OH:51])[C@@H:37]1[O:52][P:53]([OH:56])([OH:55])=[O:54])([OH:33])=[O:32])([OH:29])=[O:28])[CH3:23]参加物
実験手順
HMG-CoA reductase catalyzes the reaction of HMG-CoA with NADPH to form mevalonate, nicotinamide adenine dinucleotide phosphate (NADP) and CoA (see FIG. 1). For these assays, the cell-free extracts were prepared as described above except that the disrupted cell suspensions were centrifuged at 7,000×g instead of 15,000×g. The reaction was followed by monitoring consumption of HMG-CoA by high performance liquid chromatography (HPLC). For the assay, a reaction mixture containing (in a final volume of 0.1 ml) 0.1 M Na2H2P2O7, pH 6.5, 2 mM DTT, 1 mM NADPH, 0.4 mM HMG-CoA, and cell-free extract (the volume of extract was varied, with the balance of the 0.1 ml reaction made up with 0.1 M Na H2P2O7, pH 6.5) was incubated at 37° C. for 15 minutes. The reaction was monitored with HPLC (Luna C 18 (Phenomenex) reversed-phase column using a gradient of 0% Solvent B for 1 min. followed by 0-11% B over 42 min. Solvent A was 86:14 (v/v) 40 mM potassium phosphate (pH 6.0)/methanol. Solvent B was methanol. The wavelength for detection was 260 nm).
出典と根拠
- ORDレコード
- ord-c2798f51d74a4f999f48d5598fd24530
- 出典データセット
- uspto-grants-2010_11
- 特許番号
- US07838279B2
関連ノート
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