SHEET THE SHEET STRUCTURE OF THIS MOLECULE IS BIFURCATED. IN ORDER TO REPRESENT THIS FEATURE IN ... SHEET THE SHEET STRUCTURE OF THIS MOLECULE IS BIFURCATED. IN ORDER TO REPRESENT THIS FEATURE IN THE SHEET RECORDS BELOW, TWO SHEETS ARE DEFINED.
分子量: 71879.953 Da / 分子数: 2 / 由来タイプ: 組換発現 詳細: RESIDUES A382 AND B382 ARE ACTIVE SITE TYROSINE RESIDUES, WHICH WERE AUTOCATALYTICALLY MODIFIED TO BECOME TPQ, ARE COVALENTLY LINKED TO THE SUICIDE INHIBITOR TRANS-2-PHENYLCYCLOPROPYL-1-AMINE. ...詳細: RESIDUES A382 AND B382 ARE ACTIVE SITE TYROSINE RESIDUES, WHICH WERE AUTOCATALYTICALLY MODIFIED TO BECOME TPQ, ARE COVALENTLY LINKED TO THE SUICIDE INHIBITOR TRANS-2-PHENYLCYCLOPROPYL-1-AMINE. THIS MODIFIED TPQ IS NAMED TCQ 由来: (組換発現) ARTHROBACTER GLOBIFORMIS (バクテリア) プラスミド: PAGAO2 / 発現宿主: ESCHERICHIA COLI (大腸菌) / 株 (発現宿主): BL21(DE3) / 参照: UniProt: P46881, EC: 1.4.3.6
RESIDES 1-2, MT, ARE THOUGHT TO BE CLEAVED. THE WILD TYPE MATURE PROTEIN IS COMPOSED OF RESIDUES 3- ...RESIDES 1-2, MT, ARE THOUGHT TO BE CLEAVED. THE WILD TYPE MATURE PROTEIN IS COMPOSED OF RESIDUES 3-638. RESIDUES 639- 640, SN, IS A PRE-TAG SPACER. RESIDUES 641-648, WSHPQFEK, IS A STREP-TAG II. SEE JUDA ET AL. (2001) PROTEIN EXPRESSION AND PURIFICATION, 22, 455-461
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実験情報
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実験
実験
手法: X線回折 / 使用した結晶の数: 1
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試料調製
結晶
マシュー密度: 3 Å3/Da / 溶媒含有率: 59 %
結晶化
温度: 293 K / 手法: 蒸気拡散法, ハンギングドロップ法 / pH: 7 詳細: AGAO CRYSTALS WERE GROWN IN ABOUT 2 WEEKS BY HANGING-DROP VAPOR DIFFUSION AT 293K. THE RESERVOIR WAS 1.0-1.5M (NH4)2SO4, 2MM CUSO4, 0.15M NA CITRATE, PH 6.0-7.5. THE DROPS WERE 1.5MICROL OF ...詳細: AGAO CRYSTALS WERE GROWN IN ABOUT 2 WEEKS BY HANGING-DROP VAPOR DIFFUSION AT 293K. THE RESERVOIR WAS 1.0-1.5M (NH4)2SO4, 2MM CUSO4, 0.15M NA CITRATE, PH 6.0-7.5. THE DROPS WERE 1.5MICROL OF 11.7 MG/ML PROTEIN, 0.05M HEPES, PH 7.0, PLUS 1.5MICROL OF RESERVOIR SOLUTION. A SINGLE CRYSTAL WAS SOAKED WITH TCP BY SLOWLY ADDING TCP TO THE DROP TO ACHIEVE A 100 FOLD EXCESS OF TCP OVER ENZYME ACTIVE SITES. THE CRYSTAL WAS CRYOPROTECTED BY GRADUAL INCREMENTAL SOAKING IN RESERVOIR SOLUTION MIXED WITH GLYCEROL, WITH THE CONCENTRAION OF TCP BEING MAINTAINED CONSTANT. THE CONCENTRATION OF GLYCEROL WAS INCREASED FROM 0 TO 30% IN 2-3% INCREMENTS DURING 2 HOURS.
解像度: 1.65→28.28 Å / Cor.coef. Fo:Fc: 0.967 / Cor.coef. Fo:Fc free: 0.958 / SU B: 1.782 / SU ML: 0.058 / TLS residual ADP flag: LIKELY RESIDUAL / 交差検証法: THROUGHOUT / ESU R: 0.078 / ESU R Free: 0.08 / 立体化学のターゲット値: MAXIMUM LIKELIHOOD 詳細: HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS. TCQ WAS INITIALLY REFINED WITH MINIMAL RESTRAINTS. IT WAS DETERMINED THAT THE TCQ WAS IN THE SUBSTRATE SCHIFF BASE FORM. IN LATE REFINEMENT, ...詳細: HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS. TCQ WAS INITIALLY REFINED WITH MINIMAL RESTRAINTS. IT WAS DETERMINED THAT THE TCQ WAS IN THE SUBSTRATE SCHIFF BASE FORM. IN LATE REFINEMENT, THE TCQ WAS RESTRAINED TO THE SUBSTRATE SCHIFF BASE FORM.
Rfactor
反射数
%反射
Selection details
Rfree
0.199
10049
5 %
RANDOM
Rwork
0.171
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obs
0.172
190611
99.2 %
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溶媒の処理
イオンプローブ半径: 0.8 Å / 減衰半径: 0.8 Å / VDWプローブ半径: 1.4 Å / 溶媒モデル: BABINET MODEL WITH MASK