Thompson L F, Willis R C, Stoop J W, Seegmiller J E
Proc Natl Acad Sci U S A. 1978 Aug;75(8):3722-6. doi: 10.1073/pnas.75.8.3722.
Rates of purine synthesis de novo, as measured by the incorporation of [14C]formate into newly synthesized purines, have been determined in cultured human fibroblasts derived from normal individuals and from patients deficient in adenosine deaminase, purine nucleoside phosphorylase, or hypoxanthine phosphoribosyltransferase, three consecutive enzymes of the purine salvage pathway. All four types of cell lines are capable of incorporating [14C]formate into purines at approximately the same rate when the assays are conducted in purine-free medium. The purine overproduction that is characteristic of a deficiency in either the transferase or the phosphorylase and that results from a block in purine reutilization can be demonstrated by the resistance of [14C]formate incorporation into purines to inhibition by hypoxanthine in the case of hypoxanthine phosphoribosyltransferase-deficient fibroblasts and by resistance to inhibition by inosine in the case of purine nucleoside phosphorylase-deficient fibroblasts.
通过将[14C]甲酸掺入新合成的嘌呤中来测定从头嘌呤合成的速率,已在源自正常个体以及腺苷脱氨酶、嘌呤核苷磷酸化酶或次黄嘌呤磷酸核糖基转移酶(嘌呤补救途径的三种连续酶)缺乏症患者的培养人成纤维细胞中进行了测定。当在无嘌呤培养基中进行测定时,所有四种类型的细胞系都能够以大致相同的速率将[14C]甲酸掺入嘌呤中。在次黄嘌呤磷酸核糖基转移酶缺乏的成纤维细胞中,[14C]甲酸掺入嘌呤对次黄嘌呤抑制的抗性,以及在嘌呤核苷磷酸化酶缺乏的成纤维细胞中对肌苷抑制的抗性,可以证明由于嘌呤再利用受阻而导致的转移酶或磷酸化酶缺乏所特有的嘌呤过量产生。