Davidson B L, Tarlé S A, Van Antwerp M, Gibbs D A, Watts R W, Kelley W N, Palella T D
Department of Internal Medicine, University of Michigan, Ann Arbor 48109.
Am J Hum Genet. 1991 May;48(5):951-8.
Complete hypoxanthine-guanine phosphoribosyltransferase (HPRT) deficiency causes the Lesch-Nyhan syndrome, an X-linked, purine metabolism disorder manifested by hyperuricemia, hyperuricaciduria, and neurologic dysfunction. Partial HPRT deficiency causes hyperuricemia and gout. One requirement for understanding the molecular basis of HPRT deficiency is the determination of which amino acids in this salvage enzyme are necessary for structural or catalytic competence. In this study we have used the PCR coupled with direct sequencing to determine the nucleotide and subsequent amino acid changes in 22 subjects representing 17 unrelated kindreds from the United Kingdom. These mutations were confirmed by using either RNase mapping or Southern analyses. In addition, experiments were done to determine enzyme activity and electrophoretic mobility, and predictive paradigms were used to study the impact of these amino acid substitutions on secondary structure.
完全性次黄嘌呤 - 鸟嘌呤磷酸核糖转移酶（HPRT）缺乏会导致莱施 - 奈恩综合征，这是一种X连锁的嘌呤代谢紊乱疾病，表现为高尿酸血症、高尿酸尿症和神经功能障碍。部分HPRT缺乏会导致高尿酸血症和痛风。理解HPRT缺乏分子基础的一个必要条件是确定这种补救酶中的哪些氨基酸对于结构或催化活性是必需的。在本研究中，我们使用聚合酶链反应（PCR）结合直接测序来确定来自英国的17个无亲缘关系家系的22名受试者中的核苷酸及随后的氨基酸变化。这些突变通过核糖核酸酶图谱分析或Southern分析得以证实。此外，还进行了实验以确定酶活性和电泳迁移率，并使用预测范式来研究这些氨基酸取代对二级结构的影响。
