詳細 | タイプ | Solution-ID | 内容 | 詳細 | Label | 溶媒系 |
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solid1 | 1.5 mM [U-13C; U-15N] TPX2a5-7, 100 uM Microtubules, 25 mM phosphate buffer, 100% H2O | The TPX2 protein is fully protonated uniformly labeled with U-[13C6]-glucose and 15NH4Cl. The paclitaxel-stabilized microtubules are polymerized from bovine brain tubulin in BRB80 buffer. The mixture of TPX2 and microtubules were centrifuged through a 40% glycerol-BRB80 cushion. | protonated U-[13C,15N]-TPX2a5-7/MT assemblies | 100% H2Osolid2 | 1.5 mM [U-13C; U-15N],D; 100% back-exchanged TPX2a5-7, 100 uM Microtubules, 25 mM phosphate buffer, 100% H2O | The deuterated U-[13C,15N]-labeled TPX2a5-7 was overexpressed in minimal media in 100% D2O supplemented with 15NH4Cl and U-[13C6, D7]-glucose. The exchangeable sites in TPX2 protein was back-exchanged in 100% H2O. The paclitaxel-stabilized microtubules are polymerized from bovine brain tubulin in BRB80 buffer. The mixture of TPX2 and microtubules were centrifuged through a 40% glycerol-BRB80 cushion. | deuterated U-[13C,15N]-labeled TPX2a5-7/MT assemblies | 100% H2O | | | |
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試料 | 濃度 (mg/ml) | 構成要素 | Isotopic labeling | Solution-ID |
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1.5 mMTPX2a5-7 | [U-13C; U-15N] | 1 | 100 uMMicrotubulesnatural abundance1 | 1.5 mMTPX2a5-7 | [U-13C; U-15N],D; 100% back-exchanged | 2 | 100 uMMicrotubulesnatural abundance2 | 25 mMphosphate buffernatural abundance1 | 25 mMphosphate buffernatural abundance2 | | | | | | | | | | | | | | |
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試料状態 | Conditions-ID | イオン強度 | Label | pH | 圧 (kPa) | 温度 (K) |
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1 | 25 mM phosphate, 8 mM MgCl2, 1mM DTT, 1mM EGTA Not definedProtonated TPX2/MTs6.8 1 atm276 K2 | 25 mM phosphate, 8 mM MgCl2, 1mM DTT, 1mM EGTA Not definedDeuterated TPX2/MTs (60 kHz MAS)6.8 1 atm283 K3 | 25 mM phosphate, 8 mM MgCl2, 1mM DTT, 1mM EGTA Not definedDeuterated TPX2/MTs (fast MAS2)6.8 1 atm285 K4 | 25 mM phosphate, 8 mM MgCl2, 1mM DTT, 1mM EGTA Not definedDeuterated TPX2/MTs (110 kHz ultrafast MAS)6.8 1 atm288 K5 | 25 mM phosphate, 8 mM MgCl2, 1mM DTT, 1mM EGTA Not definedProtonated TPX2/MTs (higher temp) | | | | | | | | | | | | | | | | | | | | | |
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