正常人及Ⅵ型糖原贮积病患者溶血产物中的糖原磷酸化酶及其转换酶。磷酸化酶激酶缺乏症的研究。

Glycogen phosphorylase and its converter enzymes in haemolysates of normal human subjects and of patients with type VI glycogen-storage disease. A study of phosphorylase kinase deficiency.

作者信息

Lederer B, Van Hoof F, Van den Berghe G, Hers H

出版信息

Biochem J. 1975 Apr;147(1):23-35. doi: 10.1042/bj1470023.

DOI:10.1042/bj1470023

PMID:168880

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1165371/

Abstract

The properties of phosphorylase a, phosphorylase b, phosphorylase kinase and phosphorylase phosphatase present in a human haemolysate were investigated. The two forms of phosphorylase have the same affinity for glucose 1-phosphate but greatly differ in Vmax. Phosphorylase b is only partially stimulated by AMP, since, in the presence of the nucleotide, it is about tenfold less active than phosphorylase a. In a fresh human haemolysate phosphorylase is mostly in the b form; it is converted into phosphorylase a by incubation at 20degreesC, and this reaction is stimulated by glycogen and cyclic AMP. Once activated, the enzyme can be inactivated after filtration of the haemolysate on Sephadex G-25. This inactivation is stimulated by caffeine and glucose and inhibited by AMP and fluoride. The phosphorylase kinase present in the haemolysate can also be measured by the rate of activation of added muscle phosphorylase b, on addition of ATP and Mg2+. 2. The activity of phosphorylase kinase was measured in haemolysates obtained from a series of patients who had been classified as suffering from type VI glycogenosis. In nine patients, all boys, an almost complete deficiency of phosphorylase kinase was observed in the haemolysate and, when it could be assayed, in the liver. A residual activity, about 20% of normal, was found in the leucocyte fraction, whereas the enzyme activity was normal in the muscle. These patients suffer from the sex-linked phosphorylase kinase deficiency previously described by others. Two pairs of siblings, each time brother and sister, displayed a partial deficiency of phosphorylase kinase in the haemolysate and leucocytes and an almost complete deficiency in the liver. This is considered as being the autosomal form of phosphorylase kinase deficiency. Other patients were characterized by a low activity of total (a+b) phosphorylase and a normal or high activity of phosphorylase kinase in their haemolysate.

摘要

对人溶血产物中存在的磷酸化酶a、磷酸化酶b、磷酸化酶激酶和磷酸化酶磷酸酶的特性进行了研究。磷酸化酶的两种形式对1-磷酸葡萄糖具有相同的亲和力，但最大反应速度（Vmax）差异很大。磷酸化酶b仅受到腺苷一磷酸（AMP）的部分刺激，因为在核苷酸存在的情况下，它的活性比磷酸化酶a低约十倍。在新鲜的人溶血产物中，磷酸化酶大多为b型；在20℃孵育时可将其转化为磷酸化酶a，糖原和环磷酸腺苷（cAMP）可刺激此反应。一旦激活，在溶血产物经葡聚糖凝胶G-25过滤后，该酶可被灭活。咖啡因和葡萄糖可刺激这种灭活，而AMP和氟化物可抑制。溶血产物中存在的磷酸化酶激酶也可通过在添加三磷酸腺苷（ATP）和镁离子（Mg2+）时添加的肌肉磷酸化酶b的激活速率来测定。2. 在一系列被归类为患有VI型糖原贮积症的患者的溶血产物中测定了磷酸化酶激酶的活性。在9名患者（均为男孩）中，在溶血产物中以及在可进行测定时在肝脏中观察到几乎完全缺乏磷酸化酶激酶。在白细胞部分发现了约为正常水平20%的残余活性，而肌肉中的酶活性正常。这些患者患有先前其他人描述的X连锁磷酸化酶激酶缺乏症。两对同胞（每次均为兄妹）在溶血产物和白细胞中表现出部分磷酸化酶激酶缺乏，而在肝脏中几乎完全缺乏。这被认为是磷酸化酶激酶缺乏的常染色体形式。其他患者的特征是其溶血产物中总（a+b）磷酸化酶活性低，而磷酸化酶激酶活性正常或高。