BETA2/NeuroD protein transduction requires cell surface heparan sulfate proteoglycans.
BETA2/NeuroD protein transduction requires cell surface heparan sulfate proteoglycans.
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DOI：
10.1089/hum.2006.118
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发表时间：
2006-12
影响因子：
4.2
通讯作者：
H. Noguchi;M. Ueda;S. Matsumoto;N. Kobayashi;S. Hayashi
中科院分区：
文献类型：
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作者：
H. Noguchi;M. Ueda;S. Matsumoto;N. Kobayashi;S. Hayashi
BETA2/NeuroD蛋白对于调节胰岛素基因转录和胰岛细胞（包括胰岛素和胰高血糖素产生细胞）的终末分化是重要的。我们报道了BETA2/NeuroD蛋白可以渗透几种细胞类型，包括胰岛，因为其结构中富含精氨酸和赖氨酸的蛋白转导结构域（PTD）序列。在这里，我们提供了细胞膜硫酸乙酰肝素蛋白多糖参与细胞外BETA2/NeuroD内化的遗传和生化证据。我们测试了可溶性糖胺聚糖（GAG）是否可以抑制BETA 2/NeuroD内化。肝素几乎完全阻止BETA 2/NeuroD进入，而硫酸软骨素A、B和C仅引起有限的抑制。此外，用肝素酶III治疗损害了BETA 2/NeuroD内化，而用软骨素酶ABC或软骨素酶AC治疗在抑制BETA 2/NeuroD内化方面完全无效。我们还检查了源自CHOK 1细胞并且在GAG生物合成中有缺陷的各种突变细胞系。使用突变细胞系的观察结果支持硫酸乙酰肝素的选择性硫酸化是NeuroD/硫酸乙酰肝素识别的重要决定因素的观点。这些数据表明，细胞表面硫酸乙酰肝素蛋白聚糖是BETA 2/NeuroD内化所必需的，并且BETA 2/NeuroD蛋白转导可能是一种安全且有价值的策略，用于增强胰岛素基因转录而不需要基因转移技术。
BETA2/NeuroD protein is important for regulating insulin gene transcription and for the terminal differentiation of islet cells, including insulin- and glucagon-producing cells. We reported that BETA2/NeuroD protein can permeate several cell types, including pancreatic islets, because of an arginine- and lysine-rich protein transduction domain (PTD) sequence in its structure. Here we provide genetic and biochemical evidence that cell membrane heparan sulfate proteoglycans are involved in extracellular BETA2/NeuroD internalization. We tested whether soluble glycosaminoglycans (GAGs) could inhibit BETA2/NeuroD internalization. Heparin almost completely prevented BETA2/NeuroD entry, whereas chondroitin sulfate A, B, and C caused only limited inhibition. Moreover, treatment with heparinase III impaired BETA2/NeuroD internalization, whereas treatment with chondroitinase ABC, or with chondroitinase AC, was completely ineffective in inhibiting BETA2/NeuroD internalization. We also examined various mutant cell lines originating from CHOK1 cells and defective in GAG biosynthesis. The observation using mutant cell lines supports the notion that the selective sulfation of heparan sulfate is an important determinant for NeuroD/heparan sulfate recognition. These data indicate that cell surface heparan sulfate proteoglycans are required for BETA2/NeuroD internalization and that BETA2/NeuroD protein transduction could be a safe and valuable strategy for enhancing insulin gene transcription without requiring gene transfer technology.