High-resolution proton nuclear magnetic resonance studies of the glucocerebrosidase activator protein from Gaucher spleen.
High-resolution proton nuclear magnetic resonance studies of the glucocerebrosidase activator protein from Gaucher spleen.
复制标题
来自戈谢脾的葡萄糖脑苷脂酶激活蛋白的高分辨率质子核磁共振研究。
DOI：
10.1021/bi00344a052
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发表时间：
1985
期刊：
影响因子：
2.9
通讯作者：
Mishra,PK
中科院分区：
文献类型：
--
作者：
Sheh,L;Glew,RH;Bothner-By,AA;Mishra,PK
宾夕法尼亚州匹兹堡市，梅龙大学化学系15213接收于1985年2月1日摘要：用600 MHz质子NMR光谱法研究了从戈谢病患者脾脏中获得的热稳定蛋白因子（HSF），该因子可激活葡萄糖脑苷脂酶。一些芳香族和脂肪族的共振的自旋去耦，pH滴定，和分辨率增强实验的基础上进行了测试。高场环电流移动的脂肪族区域和低场芳香族区域进行了检查核Overhauser效应（NOE）的方法，使用脉冲傅里叶变换光谱和相关光谱。研究发现，脂肪族氨基酸残基上的一些高场位移甲基和某些亚甲基与芳香族氨基酸残基处于邻近关系，形成了一个紧密的疏水聚簇位点。特别令人感兴趣的是，酪氨酸A，苯丙氨酸A，色氨酸Bh和色氨酸B2被发现位于靠近一个簇的脂肪族残基，表明疏水位点的HSF是构象刚性和其三级结构非常紧凑。提出了HSF疏水位点的二维结构模型。陶赫尔氏病是一种遗传性溶酶体贮积病，其中由于葡糖脑苷脂酶（葡糖脑苷脂：0-葡糖苷酶; EC 3.2. 1.45）活动（Lee，1968; Fredrickson & Sloan，1972;布雷迪& Berger，1983）。近年来，许多努力致力于提高我们对这种酶的调节的理解，并解释临床极端的
Department of Chemistry, Carnegie-Mellon University, Pittsburgh, Pennsylvania 15213 Received February 1, 1985 abstract: A heat-stable protein factor (HSF) obtained from the spleen of a patient with Gaucher’s disease that activates glucocerebrosidase was studied by 600-MHz proton NMR spectroscopy. Assignments for a number of aromatic and aliphatic resonances were made on the basis of spin-decoupling, pH-titration, and resolution-enhancement experiments. The upfield ring current shifted aliphatic region and the downfield aromatic region were examined by nuclear Overhauser effect (NOE) methods using both pulsed Fourier-transform spectroscopy and correlation spectroscopy. It was found that a number of upfield-shifted methyl groups and certain methylene groups of specific aliphatic amino acid residues are in proximity relationships with several aromatic residues, forming a compact hydrophobic clustering site. Of special interest, tyrosine A, phenylalanine A, tryptophan Bh and tryptophan B2 were found to be located close to a cluster of aliphatic residues, indicating that the hydrophobic site of the HSF is conformationally rigid and its tertiary structure very compact. A two-dimensional structural model of the hydrophobic site of HSF is proposed.(jTaucher’s disease is an inherited lysosomal storage disease in which glucocerebroside accumulates in mononuclear pha-gocytic cells of the reticuloendothelial systembecause of a marked deficiency of glucocerebrosidase (glucocerebroside:0-glucosidase; EC 3.2. 1.45) activity (Lee, 1968; Fredrickson & Sloan, 1972; Brady & Barranger, 1983). In recentyears, much effort has been devoted to improving our understanding of the regulation of the enzyme and to explaining the molecular basis of the distinction between the clinical extremes of