Analysis of a rare functional truncating mutation rs61757459 in vaspin (SERPINA12) on circulating vaspin levels
Analysis of a rare functional truncating mutation rs61757459 in vaspin (SERPINA12) on circulating vaspin levels
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DOI：
10.1007/s00109-013-1062-9
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发表时间：
2013-11-01
影响因子：
4.7
通讯作者：
Kovacs, Peter
中科院分区：
文献类型：
--
作者：
Breitfeld, Jana;Heiker, John T.;Kovacs, Peter
最近的一项全基因组关联研究表明，vaspin基因内的遗传变异可能有助于循环血清内脏脂肪组织来源的丝氨酸蛋白酶抑制剂（vaspin）浓度的变化。在这里，我们分析了预测提前终止密码子的罕见变异rs61757459的功能后果及其对循环血清vaspin浓度的影响。为了确定遗传变异，我们对48名无关的高加索人进行了vaspin基因测序。随后在三个代谢特征良好的德国队列（N = 4，019）中对Rs61757459进行基因分型。我们解决了rs61757459对vaspin晶体结构的影响，并研究了其对vaspin表达的影响，在体内以及体外使用各种细胞系（大肠杆菌，HEK 293）。沿着先前报道的常见遗传变异，vaspin的测序揭示了一种罕见的变异（rs61757459;次要等位基因频率：1%），其预测提前终止密码子p.R211X。与非携带者相比，这种突变的杂合子携带者循环vaspin水平较低。截短的vaspin，估计为24.5 kDa，在电脑结构分析表明，由于缺乏核心结构域的错误折叠和潜在的不稳定性。事实上，在E.大肠杆菌和裂解液中，但在HEK 293细胞的上清液中不存在。我们得出结论，rs61757459是一个功能突变，导致截短的蛋白质，其不稳定性可能导致血清vaspin水平降低。（rs61757459）在vaspin中编码终止密码子p.R211X与较低的循环vaspin浓度有关。结构分析表明由于缺乏核心结构域而导致错误折叠和不稳定。在E.大肠杆菌和裂解液中，但在HEK 293细胞的上清液中不存在。
A recent genome-wide association study suggests that genetic variation within the vaspin gene might contribute to the variability in circulating serum visceral adipose tissue-derived serine protease inhibitor (vaspin) concentrations. Here, we analyzed the functional consequences of the rare variant rs61757459 predicting a premature stop codon and its impact on circulating serum vaspin concentrations. In order to identify genetic variation, we sequenced the vaspin gene in 48 nonrelated Caucasian subjects. Rs61757459 was subsequently genotyped in three metabolically well-characterized German cohorts (N = 4,019). We addressed the impact of rs61757459 on the crystal structure of vaspin and investigated its effects on vaspin expression in vivo as well as in vitro using various cell lines (Escherichia coli, HEK293). Along with previously reported common genetic variants, sequencing of vaspin revealed a rare variant (rs61757459; minor allele frequency: 1 %) which predicts a premature stop codon p.R211X. Heterozygous carriers of this mutation had lower circulating vaspin levels when compared with noncarriers. In silico structure analysis of the truncated vaspin, which was estimated to be 24.5 kDa, suggested misfolding and potential instability due to the absence of core structural domains. Indeed, the truncated protein was detected after recombinant expression in E. coli and in lysate, but not in supernatant of HEK293 cells. We conclude that rs61757459 is a functional mutation that results in a truncated protein whose instability likely results in reduced serum vaspin levels.A rare variant (rs61757459) in vaspin coding for the stop codon p.R211X is related to lower circulating vaspin concentrations.Structure analysis suggests misfolding and instability due to the absence of core structural domains.The truncated protein is detectable after recombinant expression in E. coli and in lysate, but not in supernatant of HEK293 cells.