Bose Neelanjan, Zee Tiffany, Kapahi Pankaj, Stoller Marshall L
Department of Urology, University of California, San Francisco, CA, USA.
Buck Institute for Research on Aging, Novato, CA, USA.
Bio Protoc. 2017 Jul 20;7(14). doi: 10.21769/BioProtoc.2417.
Cystinuria is a rare genetic disorder characterized by recurrent, painful kidney stones, primarily composed of cystine, the dimer of the amino acid cysteine (Sumorok and Goldfarb, 2013). Using a mouse model of cystinuria, we have recently shown that administration of drugs that increase cystine solubility in the urine can be a novel therapeutic strategy for the clinical management of the disease (Zee ., 2017). There is a large unmet need in the field for developing new drugs for cystinuria. To that end, here we describe a simple cystine solubility assay that is amenable for screening compounds to identify potential drugs that may influence cystine solubility. The assay includes preparing a supersaturated solution of cystine, incubating this solution with drug(s) of choice, and finally using high pressure liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) to quantify the amount of cystine precipitated under various conditions.
胱氨酸尿症是一种罕见的遗传性疾病，其特征是反复出现疼痛性肾结石，主要由胱氨酸组成，胱氨酸是氨基酸半胱氨酸的二聚体（苏莫罗克和戈德法布，2013年）。我们最近利用胱氨酸尿症小鼠模型表明，给予可增加尿液中胱氨酸溶解度的药物可能是该疾病临床管理的一种新治疗策略（泽等人，2017年）。在开发用于胱氨酸尿症的新药方面，该领域存在巨大的未满足需求。为此，我们在此描述一种简单的胱氨酸溶解度测定法，该方法适用于筛选化合物，以鉴定可能影响胱氨酸溶解度的潜在药物。该测定法包括制备胱氨酸的过饱和溶液，将该溶液与所选药物一起孵育，最后使用高压液相色谱 - 串联质谱法（HPLC-MS/MS）来定量在各种条件下沉淀的胱氨酸量。
