Brophy Megan L, Murphy John E, Bell Robert D
Rare Disease Research Unit, Worldwide Research, Development and Medicine, Pfizer, Inc. Cambridge, MA 02139, USA.
J Biol Methods. 2021 Jun 30;8(2):e149. doi: 10.14440/jbm.2021.355. eCollection 2021.
Galactosemias are a family of autosomal recessive genetic disorders resulting from impaired enzymes of the Leloir pathway of galactose metabolism including galactokinase, galactose uridyltransferase, and UDP-galactose 4-epimerase that are critical for conversion of galactose into glucose-6-phosphate. To better understand pathophysiological mechanisms involved in galactosemia and develop novel therapies to address the unmet need in patients, it is important to develop reliable assays to measure the activity of the Leloir pathway enzymes. Here we describe in-depth methods for indirectly measuring galacose-1-phosphate uridyltransferase activity in cell culture and animal tissues.
半乳糖血症是一类常染色体隐性遗传疾病，由半乳糖代谢的Leloir途径中的酶受损所致，这些酶包括半乳糖激酶、半乳糖尿苷转移酶和UDP - 半乳糖4 - 表异构酶，它们对于将半乳糖转化为6 - 磷酸葡萄糖至关重要。为了更好地理解半乳糖血症所涉及的病理生理机制，并开发新的疗法以满足患者未被满足的需求，开发可靠的检测方法来测量Leloir途径酶的活性非常重要。在此，我们描述了在细胞培养物和动物组织中间接测量1 - 磷酸半乳糖尿苷转移酶活性的深入方法。
