Glycosylation and processing of pro-B-type natriuretic peptide in cardiomyocytes.
Glycosylation and processing of pro-B-type natriuretic peptide in cardiomyocytes.
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DOI：
10.1016/j.bbrc.2011.06.192
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发表时间：
2011-08-05
影响因子：
3.1
通讯作者：
Wu, Qingyu
中科院分区：
文献类型：
--
作者：
Peng, Jianhao;Jiang, Jingjing;Wang, Wei;Qi, Xiaofei;Sun, Xue-Long;Wu, Qingyu
B型利钠肽（BNP）及其相关肽是诊断心力衰竭的生物标志物。最近的研究确定了几个O-糖基化位点，包括Thr-71，在人pro-BNP上，但功能意义尚不清楚。在这项研究中，我们分析了前BNP在心肌细胞中的糖基化和蛋白水解加工。人pro-BNP野生型（WT）和突变体在HEK 293细胞和鼠HL-1心肌细胞中表达。采用免疫沉淀法和Western blotting法检测Pro-BNP和BNP。使用糖基化酶和糖基化抑制剂来检查pro-BNP上的碳水化合物。还检测了弗林蛋白酶和corin表达对细胞中pro-BNP加工的影响。我们发现在HEK 293细胞中，重组pro-BNP含有大量的具有末端寡唾液酸的O-聚糖。Thr-71突变减少了pro-BNP上的O-聚糖，并增加了pro-BNP加工。在HL-1心肌细胞中，残基Thr-71几乎不含O-聚糖，pro-BNP WT和T71 A突变体的处理类似。在HEK 293细胞中，pro-BNP被弗林蛋白酶加工。Arg-73和Arg-76的突变，而不是Lys-79，阻止了pro-BNP的加工。在表达弗林蛋白酶和corin的HL-1心肌细胞中，Arg-73、Arg-76和Lys-79的单突变或双突变并不能阻止pro-BNP的加工。只有当所有这三个残基都发生突变时，pro-BNP加工才被完全阻断。我们的数据表明，pro-BNP在心肌细胞中的糖基化显着不同的HEK 293细胞。在HEK 293细胞中，弗林蛋白酶在Arg-76处切割pro-BNP，而在心肌细胞中，corin在包括Arg-73、Arg-76和Lys-79的多个残基处切割pro-BNP。
B-type natriuretic peptide (BNP) and its related peptides are biomarkers for the diagnosis of heart failure. Recent studies identified several O-glycosylation sites, including Thr-71, on human pro-BNP but the functional significance was unclear. In this study, we analyzed glycosylation and proteolytic processing of pro-BNP in cardiomyocytes. Human pro-BNP wild-type (WT) and mutants were expressed in HEK 293 cells and murine HL-1 cardiomyocytes. Pro-BNP and BNP were analyzed by immunoprecipitation and Western blotting. Glycosidases and glycosylation inhibitors were used to examine carbohydrates on pro-BNP. The effects of furin and corin expression on pro-BNP processing in cells also were examined. We found that in HEK 293 cells, recombinant pro-BNP contained significant amounts of O-glycans with terminal oligosialic acids. Mutation at Thr-71 reduced O-glycans on pro-BNP and increased pro-BNP processing. In HL-1 cardiomyocytes, residue Thr-71 contained little O-glycans, and pro-BNP WT and T71A mutant were processed similarly. In HEK 293 cells, pro-BNP was processed by furin. Mutations at Arg-73 and Arg-76, but not Lys-79, prevented pro-BNP processing. In HL-1 cardiomyocytes, which express furin and corin, single or double mutations at Arg-73, Arg-76 and Lys-79 did not prevent pro-BNP processing. Only when all these three residues were mutated, was pro-BNP processing completely blocked. Our data indicate that pro-BNP glycosylation in cardiomyocytes differed significantly from that in HEK 293 cells. In HEK 293 cells, furin cleaved pro-BNP at Arg-76 whereas in cardiomyocytes corin cleaved pro-BNP at multiple residues including Arg-73, Arg-76 and Lys-79.
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影响因子：
9.3
作者：
Jiang J;Pristera N;Wang W;Zhang X;Wu Q
通讯作者：
Wu Q
DOI：
10.1152/ajpregu.00569.2006
发表时间：
2007-02-01
影响因子：
2.8
作者：
Boerrigter, Guido;Costello-Boerrigter, Lisa C.;Burnett, John C., Jr.
通讯作者：
Burnett, John C., Jr.
影响因子：
158.5
作者：
Maisel, AS;Krishnaswamy, P;McCullough, PA
通讯作者：
McCullough, PA
DOI：
10.1152/ajpheart.00399.2010
发表时间：
2010-11-01
影响因子：
4.8
作者：
Chen, Shenghan;Sen, Subha;Wu, Qingyu
通讯作者：
Wu, Qingyu
影响因子：
3.9
作者：
Crimmins, Dan L.;Kao, Jeffrey L. -F.
通讯作者：
Kao, Jeffrey L. -F.