5'-Atp → Palmitic Acid

HRID 1278911

반응식

HRID 1278911 반응식
C(N(CC(O)=O)CC(O)=O)CN(CC(O)=O)CC(O)=O.CC(C[C@H](NC(C)=O)C(N[C@H](C(N[C@H:37]([C:45]([OH:47])=[O:46])[CH2:38][CH2:39][CH2:40]N=C(N)N)=O)CC(C)C)=O)C.CCC(CO[C:59](C(N(CC[NH+](C)C)C)=O)([C:66]1[CH:71]=[CH:70][CH:69]=[CH:68][CH:67]=1)[C:60]1C=C[CH:63]=[CH:62][CH:61]=1)CC.[Cl-].[C@@H]1(N2C3N=CN=C(N)C=3N=C2)O[C@H](COP(OP(OCC([C@H](C(NCCC(NCCS)=O)=O)O)(C)C)(O)=O)(O)=O)[C@@H](OP(O)(O)=O)[C@H]1O.P(OC[C@H]1O[C@@H](N2C3N=CN=C(N)C=3N=C2)[C@H](O)[C@@H]1O)(OP(OP(O)(O)=O)(O)=O)(=O)O.CCCCCCCCCCCCOS([O-])(=O)=O.[Na+]>C1N(CCO)CCN(CCS(O)(=O)=O)C1.C(O)C(N)(CO)CO.Cl>[C:45]([OH:47])(=[O:46])[CH2:37][CH2:38][CH2:39][CH2:40][CH2:71][CH2:70][CH2:69][CH2:68][CH2:67][CH2:66][CH2:59][CH2:60][CH2:61][CH2:62][CH3:63]

참가 물질

9

반응물 6

HCID 59575'-AtpHCID 6049Ethylenediaminetetraacetic Acid2회HCID 19153X 100HCID 87642Coenzyme AHCID 189069Leupeptin acid2회HCID 3423265Sodium Lauryl Sulfate

생성물 1

HCID 985Palmitic Acid

시약·촉매·용매 2

HCID 238314-(2-Hydroxyethyl)-1-piperazineethanesulfonic acid용매HCID 93573Tromethamine hydrochloride용매2회

실험 절차

Soluble Shh was labeled with 3H-palmitic acid in a cell-free system using a modified version of a published procedure (24). A crude microsomal fraction from rat liver was prepared by subjecting a liver homogenate to sequential centrifugation at 3000×g for 10 min, 9000×g for 20 min, and 100,000×g for 30 min. The 100,000×g pellet was suspended in 10 mM HEPES pH 7.4, 10% sucrose and again centrifuged at 100,000×g for 20 min. The final pellet (derived from 10 g of liver) was suspended in 3 mL of 20 mM Tris-HCl pH 7.4, 150 mM NaCl, 1 mM EDTA, 10 μg/mL leupeptin, 0.15% Triton X-100, aliquoted, and stored at −70° C. Reactions containing 3 μg Shh, 1 μL of rat microsomes, 50 ng/mL Coenzyme A (Sigma), 0.3 mM ATP, 20 mM Tris-HCl pH 7.4, 150 mM NaCl, 1 mM EDTA, 10 μg/mL leupeptin, and 0.5 μCi-[9,10-3H]-palmitic acid (50 Ci/mmol; New England Nuclear) were performed at room temperature for 1 h. Reactions were stopped with reducing electrophoresis sample buffer, subjected to SDS-PAGE on a 10-20% gradient gel, and visualized by fluorography.

후처리

  1. customA crude microsomal fraction from rat liver was prepared
  2. wait9000×g for 20 min
  3. wait100,000×g for 30 min
  4. waitagain centrifuged at 100,000×g for 20 min
  5. customstored at −70° C
  6. waitwere performed at room temperature for 1 h

출처 및 근거

ORD 기록
ord-ca440bf402f441fcbcfc7df96221cafd
출처 데이터셋
uspto-grants-2002_09
특허 번호
US06444793B1

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