Allele-specific characterization of alanine: glyoxylate aminotransferase variants associated with primary hyperoxaluria.
Allele-specific characterization of alanine: glyoxylate aminotransferase variants associated with primary hyperoxaluria.
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DOI：
10.1371/journal.pone.0094338
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Tucker CL
中科院分区：
文献类型：
--
作者：
Lage MD;Pittman AM;Roncador A;Cellini B;Tucker CL
原发性高草酸尿1型（PH1）是一种罕见的常染色体隐性肾结石疾病，由参与乙醛酸解毒的过氧化物酶丙氨酸：乙醛酸氨基转移酶（AGT）缺乏引起。已经发现超过75种不同的AGT错义突变与PH1相关。虽然已经发现一些突变会影响酶的活性、稳定性和/或定位，但大约一半的突变是完全没有特征的。在这项研究中，我们试图系统地表征与PH1相关的AGT错义突变。为了便于分析，我们使用了两种基于酵母的高通量检测：一种用于评估AGT特异性活性，另一种用于评估蛋白质稳定性。大约30%的ph1相关错义突变与一个较小的等位基因多态性变异一起被发现，它们可以相互作用，引发对蛋白质稳定性和运输的复杂影响。为了更好地理解这种等位基因相互作用，我们在主要（野生型）和次要等位基因背景上对34个突变体进行了功能表征，确定了与次要等位基因协同作用的突变。我们根据不同等位基因的活性/稳定性结果将这些突变体分为四种不同的类别。与主等位基因背景相比，发现有12个突变体与小等位基因结合时表现出较低的活性。当在AGT二聚体结构上进行映射时，这些突变体揭示了蛋白质的局部区域，这些区域似乎对与次要等位基因变体的相互作用特别敏感。虽然对次要等位基因活性的大多数有害影响可归因于影响蛋白质稳定性的协同相互作用，但我们发现了一个突变E274D，当与次要等位基因结合时，它似乎特别影响活性。
Primary Hyperoxaluria Type 1 (PH1) is a rare autosomal recessive kidney stone disease caused by deficiency of the peroxisomal enzyme alanine: glyoxylate aminotransferase (AGT), which is involved in glyoxylate detoxification. Over 75 different missense mutations in AGT have been found associated with PH1. While some of the mutations have been found to affect enzyme activity, stability, and/or localization, approximately half of these mutations are completely uncharacterized. In this study, we sought to systematically characterize AGT missense mutations associated with PH1. To facilitate analysis, we used two high-throughput yeast-based assays: one that assesses AGT specific activity, and one that assesses protein stability. Approximately 30% of PH1-associated missense mutations are found in conjunction with a minor allele polymorphic variant, which can interact to elicit complex effects on protein stability and trafficking. To better understand this allele interaction, we functionally characterized each of 34 mutants on both the major (wild-type) and minor allele backgrounds, identifying mutations that synergize with the minor allele. We classify these mutants into four distinct categories depending on activity/stability results in the different alleles. Twelve mutants were found to display reduced activity in combination with the minor allele, compared with the major allele background. When mapped on the AGT dimer structure, these mutants reveal localized regions of the protein that appear particularly sensitive to interactions with the minor allele variant. While the majority of the deleterious effects on activity in the minor allele can be attributed to synergistic interaction affecting protein stability, we identify one mutation, E274D, that appears to specifically affect activity when in combination with the minor allele.
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影响因子：
2.6
作者：
Schlösser, T;Gätgens, C;Stahmann, KP
通讯作者：
Stahmann, KP
DOI：
10.1073/pnas.0908565107
发表时间：
2010-02-16
影响因子：
11.1
作者：
Cellini, Barbara;Montioli, Riccardo;Voltattorni, Carla Borri
通讯作者：
Voltattorni, Carla Borri
影响因子：
4.8
作者：
Hopper, Erin D.;Pittman, Adrianne M. C.;Tucker, Chandra L.
通讯作者：
Tucker, Chandra L.
影响因子：
3.3
作者：
Pittman, Adrianne M. C.;Lage, Melissa D.;Tucker, Chandra L.
通讯作者：
Tucker, Chandra L.
影响因子：
4.1
作者：
Lumb, MJ;Birdsey, GM;Danpure, CJ
通讯作者：
Danpure, CJ