Brophy P J, Vance D E
Biochem J. 1975 Dec;152(3):495-501. doi: 10.1042/bj1520495.
Elongation of fatty acids by microsomal fractions obtained from rat brain was measured by the incorporation of [2-14C]malonyl-CoA into fatty in the presence of palmitoyl-CoA or stearoyl-CoA. 2. Soluble and microsomal fractions were prepared from 21-day-old rats; density gradient centrifugation demonstrated that the stearoyl-CoA elongation system was localized in the microsomal fraction whereas fatty acid biosynthesis de novo from acetyl-CoA occurred in the soluble fraction. The residual activity de novo in the microsomal fraction was attributed to minor contamination by the soluble fraction. 3. The optimum concentration of [2-14C]malonyl-CoA for elongation of fatty acids was 25 mum for palmitoyl-CoA or stearoyl-CoA, and the corresponding optimum concentrations for the two primer acyl-CoA esters were 8.0 and 7.2 muM respectively. 4. Nadph was the preferred cofactor for fatty acid formation from palmitoyl-CoA or stearoyl-CoA, although NADH could partially replace it. 5. The stearoyl-CoA elongation system required a potassium phosphate buffer concentration of 0.075M for maximum activity; CoA (1 MUM) inhibited this elongation system by approx. 30%. 6. The fatty acids formed from malonyl-CoA and palmitoyl-CoA had a predominant chain length of C18 whereas stearoyl-CoA elongation resulted in an even distribution of fatty acids with chain lengths of C20, C22 and C24. 7. The products of stearoyl-CoA elongation were identified as primarily unesterified fatty acids. 8. The developmental pattern of fatty acid biosynthesis by rat brain microsomal preparations was studied and both the palmitoyl-CoA and stearoyl-CoA elongation systems showed large increases in activity between days 10 and 18 after birth.
通过在棕榈酰辅酶A或硬脂酰辅酶A存在的情况下,将[2-14C]丙二酰辅酶A掺入脂肪酸中,来测定从大鼠脑获得的微粒体部分对脂肪酸的延长作用。2. 从21日龄大鼠制备可溶性和微粒体部分;密度梯度离心表明,硬脂酰辅酶A延长系统定位于微粒体部分,而从乙酰辅酶A从头合成脂肪酸则发生在可溶性部分。微粒体部分中从头合成的残留活性归因于可溶性部分的少量污染。3. 对于棕榈酰辅酶A或硬脂酰辅酶A,脂肪酸延长的[2-14C]丙二酰辅酶A的最佳浓度为25μM,两种引物酰基辅酶A酯的相应最佳浓度分别为8.0和7.2μM。4. NADPH是从棕榈酰辅酶A或硬脂酰辅酶A形成脂肪酸的首选辅因子,尽管NADH可以部分替代它。5. 硬脂酰辅酶A延长系统需要0.075M的磷酸钾缓冲液浓度才能达到最大活性;辅酶A(1μM)对该延长系统的抑制作用约为30%。6. 由丙二酰辅酶A和棕榈酰辅酶A形成的脂肪酸的主要链长为C18,而硬脂酰辅酶A延长导致链长为C20、C22和C24的脂肪酸均匀分布。7. 硬脂酰辅酶A延长的产物主要鉴定为未酯化的脂肪酸。8. 研究了大鼠脑微粒体制剂中脂肪酸生物合成的发育模式,棕榈酰辅酶A和硬脂酰辅酶A延长系统在出生后第10天至18天之间的活性均大幅增加。