Casazza J P, Veech R L
J Biol Chem. 1986 Jan 15;261(2):690-8.
Equilibrium constants for reactions catalyzed by ribulose-5-phosphate 3-epimerase, [sigma xylulose-5-P]/[sigma ribulose-5-P] = 1.82, ribose-5-phosphate isomerase, [sigma Rib-5-P]/[sigma ribulose-5-P] = 1.20, transaldolase, [sigma erythrose-4-P] [sigma Fru-6-P]/[sigma sedoheptulose-7-P] [sigma glyceraldehyde 3-P] = 0.37, and transketolase, [sigma Fru-6-P] [sigma glyceraldehyde 3-P]/[sigma erythrose-4-P] [sigma xylulose-5-P] = 29.7 and [sigma Rib-5-P] [sigma xylulose-5-P]/[sigma sedoheptulose-7-P] [sigma glyceraldehyde 3-P] = 0.48, were redetermined under physiological conditions. The equilibrium constant for the combined glucose-6-P dehydrogenase and 6-phosphoglucono-gamma-lactonase reaction, [6-phosphogluconate3-] [NADPH] [H+]2/[Glc-6-P2-] [NADP+], was found to be at least 1 X 10(-9). Using these redetermined equilibrium constants, calculated values of pentose cycle intermediates, based on near equilibrium assumptions and the tissue content of Fru-6-P and glyceraldehyde 3-P, were found to be in good agreement with measured values for male Wistar rats injected with saline, 20 mumol/g pyruvate, 20 mumol/g gluconate, and 20 mumol/g ribose. Measured and calculated values for pentose cycle intermediates in saline injected animals were ribulose-5-P; 3.8 +/- 0.4 and 2.4 +/- 0.1 nmol/g; xylulose-5-P, 5.9 +/- 0.6 nmol/g and 4.3 +/- 0.2 nmol/g; sedoheptulose-7-P, 41.5 +/- 2.4 and 37.6 +/- 2.9 nmol/g; and combined sedopheptulose-7-P and Rib-5-P, 43.0 +/- 2.8 nmol/g and 40.5 +/- 3.0 nmol/g; liver content of erythrose-4-P was less than the detection limits of the assay, 2 nmol/g. Calculated erythrose-4-P was 0.23 +/- 0.01 nmol/g. Liver content of 6-phosphogluconate was 8.5 +/- 0.7 nmol/g. The free cytosolic [NADP+]/[NADPH] ratio calculated from the 6-phosphogluconate dehydrogenase redox couple, 0.0030 +/- 0.0002, was also in good agreement with that calculated from the malic enzyme redox couple, 0.0051 +/- 0.0007, and the isocitrate dehydrogenase redox couple, 0.0066 +/- 0.0008. These data indicate the interdependence of the liver content of glycolytic intermediates and pentose cycle intermediates in ad libitum fed rats.
在生理条件下重新测定了由磷酸核糖 - 5 - 磷酸3 - 表异构酶催化反应的平衡常数,即[木酮糖 - 5 - 磷酸]/[核糖 - 5 - 磷酸] = 1.82;核糖 - 5 - 磷酸异构酶的平衡常数,即[核糖 - 5 - 磷酸]/[核糖 - 5 - 磷酸] = 1.20;转醛醇酶的平衡常数,即[赤藓糖 - 4 - 磷酸][果糖 - 6 - 磷酸]/[景天庚酮糖 - 7 - 磷酸][甘油醛 - 3 - 磷酸] = 0.37;以及转酮醇酶的平衡常数,即[果糖 - 6 - 磷酸][甘油醛 - 3 - 磷酸]/[赤藓糖 - 4 - 磷酸][木酮糖 - 5 - 磷酸] = 29.7,[核糖 - 5 - 磷酸][木酮糖 - 5 - 磷酸]/[景天庚酮糖 - 7 - 磷酸][甘油醛 - 3 - 磷酸] = 0.48。发现葡萄糖 - 6 - 磷酸脱氢酶和6 - 磷酸葡萄糖酸 - γ - 内酯酶联合反应的平衡常数,即[6 - 磷酸葡萄糖酸³⁻][NADPH][H⁺]²/[葡萄糖 - 6 - 磷酸²⁻][NADP⁺]至少为1×10⁻⁹。使用这些重新测定的平衡常数,基于接近平衡的假设以及果糖 - 6 - 磷酸和甘油醛 - 3 - 磷酸的组织含量计算得到的戊糖循环中间体的值,与注射生理盐水、20 μmol/g丙酮酸、20 μmol/g葡萄糖酸和20 μmol/g核糖的雄性Wistar大鼠的测量值高度一致。注射生理盐水动物中戊糖循环中间体的测量值和计算值分别为:磷酸核糖 - 5 - 磷酸,3.8±0.4和2.4±0.1 nmol/g;木酮糖 - 5 - 磷酸,5.9±0.6 nmol/g和4.3±0.2 nmol/g;景天庚酮糖 - 7 - 磷酸,41.5±2.4和37.6±2.9 nmol/g;景天庚酮糖 - 7 - 磷酸和核糖 - 5 - 磷酸总和,43.0±2.8 nmol/g和40.5±3.0 nmol/g;肝脏中赤藓糖 - 4 - 磷酸含量低于检测限2 nmol/g。计算得到的赤藓糖 - 4 - 磷酸为0.23±0.01 nmol/g。肝脏中6 - 磷酸葡萄糖酸含量为8.5±0.7 nmol/g。由6 - 磷酸葡萄糖酸脱氢酶氧化还原对计算得到的游离胞质[NADP⁺]/[NADPH]比值为0.0030±0.0002,与由苹果酸酶氧化还原对计算得到的0.0051±0.0007以及异柠檬酸脱氢酶氧化还原对计算得到的0.0066±0.0008也高度一致。这些数据表明在随意进食的大鼠中,糖酵解中间体和戊糖循环中间体的肝脏含量相互依赖。