Effect of sialylated O-glycans in pro-brain natriuretic peptide stability.
Effect of sialylated O-glycans in pro-brain natriuretic peptide stability.
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DOI：
10.1373/clinchem.2009.140558
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发表时间：
2010-06
影响因子：
9.3
通讯作者：
Wu Q
中科院分区：
文献类型：
--
作者：
Jiang J;Pristera N;Wang W;Zhang X;Wu Q
心房、脑和C型利钠肽（ANP、BNP和CNP）在调节多种心血管和细胞功能中很重要。在细胞中，这些肽被制成前体形式，转化为成熟形式。BNP及其相关肽是诊断心力衰竭的生物标志物。在这项研究中，我们研究了pro-ANP，pro-BNP和pro-CNP中的糖基化，这可能会改变它们的生化和代谢特性。人pro-ANP、pro-BNP和pro-CNP在人胚肾（HEK）293细胞和小鼠HL-1心肌细胞中表达，并通过免疫沉淀和Western印迹进行分析。使用去糖基化酶来测定这些肽上的碳水化合物含量。还检查了抑制0-糖基化对肽的细胞表达和稳定性的影响。在HEK 293和HL-1细胞中，来自培养基的pro-BNP而不是pro-ANP和pro-CNP具有比来自细胞裂解物的更大的分子量。用PNGase F、O-糖苷酶和唾液酸酶A消化表明pro-BNP含有O-聚糖，但不含N-聚糖。pro-BNP上的O-聚糖在其末端具有唾液酸，保护其免受O-糖苷酶消化。相比之下，pro-ANP和pro-CNP不含可检测量的N-或O-聚糖。对pro-BNP的O-糖基化的抑制并不能阻止其在细胞中的表达。然而，部分O-糖基化pro-BNP的稳定性远低于完全O-糖基化pro-BNP。O-糖基化对pro-BNP的表达不是必需的，但对其稳定性很重要。
Atrial-, brain- and C-type natriuretic peptides (ANP, BNP, and CNP) are important in regulating a variety of cardiovascular and cellular functions. In cells, these peptides are made as pro-forms that are converted to mature forms. BNP and its related peptides are biomarkers for the diagnosis of heart failure. In this study, we examined glycosylation in pro-ANP, pro-BNP and pro-CNP, which may alter their biochemical and metabolic properties. Human pro-ANP, pro-BNP, and pro-CNP were expressed in human embryonic kidney (HEK) 293 cells and murine HL-1 cardiomyocytes, and analyzed by immunoprecipitation and Western blotting. Deglycosylation enzymes were used to determine the carbohydrate content on these peptides. The effects of inhibiting O-glycosylation on cellular expression and stability of the peptides also were examined. In HEK 293 and HL-1 cells, pro-BNP, but not pro-ANP and pro-CNP, from the culture medium had a greater molecular mass than that from cell lysate. Digestion with PNGase F, O-glycosidase and sialidase A indicated that pro-BNP contained O-glycans but not N-glycans. The O-glycans on pro-BNP had sialic acids at their termini, protecting it from O-glycosidase digestion. In contrast, pro-ANP and pro-CNP contained no detectable amounts of N- or O-glycans. Inhibition of O-glycosylation on pro-BNP did not prevent its expression in the cells. However, partially O-glycosylated pro-BNP was much less stable than fully O-glycosylated pro-BNP. O-glycosylation is not necessary for pro-BNP expression but important for its stability.
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DOI：
10.1016/s0735-1097(03)00787-3
发表时间：
2003-08-20
影响因子：
24
作者：
Lainchbury, JG;Campbell, E;Richards, AM
通讯作者：
Richards, AM
影响因子：
158.5
作者：
Maisel, AS;Krishnaswamy, P;McCullough, PA
通讯作者：
McCullough, PA
影响因子：
9.3
作者：
Giuliani, Isabelle;Rieunier, Francois;Villard, Sylvie
通讯作者：
Villard, Sylvie
影响因子：
3.9
作者：
Crimmins, Dan L.;Kao, Jeffrey L. -F.
通讯作者：
Kao, Jeffrey L. -F.
影响因子：
4.8
作者：
Liao, Xudong;Wang, Wei;Wu, Qingyu
通讯作者：
Wu, Qingyu