Boza J J, Jahoor F, Reeds P J
U.S. Department of Agriculture-Agricultural Research Service, Children's Nutrition Research Center, Houston, TX, USA.
J Nutr. 1996 Jul;126(7):1749-58. doi: 10.1093/jn/126.7.1749.
The contributions of dietary nucleotides and nucleotides synthesized de novo to ribonucleic acid synthesis in vivo were estimated by feeding, from d 13 to 18 of gestation, two groups of five pregnant mice a defined diet that contained either uniformly [U13C]-labeled nucleotides or [U13C]-algal amino acids isolated from algal biomass. Ribonucleic acid and protein were isolated from mucosa, liver and fetus. Nucleosides and amino acids were isolated and converted to their trimethylsilyl and n-propyl ester, heptaflurobutyramide derivatives, respectively. The isotopic enrichments of all isotopomers were determined by gas chromatography-mass spectrometry. In the mice that ingested [U13C]-nucleotides, the isotopic enrichment of [Ul3C]-purines (0.03-0.2 mol/100 mol) was significantly (P < 0.001) less than that of [U13C]-uridine (1.5-4.2 mol/100 mol). [13C5]-Purines (0.1-0.8 mol/100 mol) and [13C4]-uridine (0.2-0.5 mol/100 mol) were detected, showing that some dietary bases and ribose were incorporated via the salvage pathway. In mice that Ingested U13C-amino acids, the isotopic enrichment (2-4.6 mol/100 mol) of the [13C2]-purines, which derive from [Ul3C]-glycine, was between 73 (liver) and 113% (fetus) of protein-bound 13C2-glycine. The isotopic enrichment (0.8-1.6 mol/100 mol) of [13C3]-uridine, an isotopomer that derives from [U13C]-aspartate, was 50 (liver) to 126% (mucosa) of [13C4]-protein-bound aspartate. The results suggest that a large majority of the bases incorporated into maternal and fetal ribonucleic acids derive from synthesis de novo.
通过在妊娠第13至18天给两组各五只怀孕小鼠喂食特定饮食,来估计膳食核苷酸和从头合成的核苷酸对体内核糖核酸合成的贡献。该特定饮食包含均匀标记的[U13C]核苷酸或从藻类生物质中分离出的[U13C]藻类氨基酸。从黏膜、肝脏和胎儿中分离出核糖核酸和蛋白质。分离出核苷和氨基酸,并分别将它们转化为三甲基硅烷基酯和正丙基酯、七氟丁酰胺衍生物。通过气相色谱 - 质谱法测定所有异构体的同位素丰度。在摄入[U13C]核苷酸的小鼠中,[Ul3C]嘌呤的同位素丰度(0.03 - 0.2摩尔/100摩尔)显著低于(P < 0.001)[U13C]尿苷的同位素丰度(1.5 - 4.2摩尔/100摩尔)。检测到[13C5]嘌呤(0.1 - 0.8摩尔/100摩尔)和[13C4]尿苷(0.2 - 0.5摩尔/100摩尔),表明一些膳食碱基和核糖通过补救途径掺入。在摄入U13C氨基酸的小鼠中,源自[Ul3C]甘氨酸的[13C2]嘌呤的同位素丰度(2 - 4.6摩尔/100摩尔)为蛋白质结合的13C2甘氨酸的73%(肝脏)至113%(胎儿)。源自[U13C]天冬氨酸的异构体[13C3]尿苷的同位素丰度(0.8 - 1.6摩尔/100摩尔)为[13C4]蛋白质结合天冬氨酸的50%(肝脏)至126%(黏膜)。结果表明,掺入母体和胎儿核糖核酸中的大部分碱基源自从头合成。