PICK1中N端酸性区域对其PDZ结构域脂质结合能力的调节

杨廷, 贾原, 肖虹, 石亚伟

中国生物化学与分子生物学报 ›› 2012, Vol. 28 ›› Issue (10) : 940-945.

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中国生物化学与分子生物学报 ›› 2012, Vol. 28 ›› Issue (10) : 940-945.
研究论文

PICK1中N端酸性区域对其PDZ结构域脂质结合能力的调节

  • 杨廷1),贾原2),肖虹3),石亚伟1)*
作者信息 +

Regulation of Liposome Binding of PDZ Domain by Its Adjacent N-terminal Acidic Region

  • YANG Ting1), JIA Yuan2), XIAO Hong 3), SHI Ya-Wei1)*
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文章历史 +

摘要

蛋白激酶Cα相互作用蛋白1(protein interacting with Cα kinase 1, PICK1)是衔接膜上受体和蛋白激酶Cα的重要蛋白.利用荧光光谱结合定点突变技术 、蛋白与脂质覆盖法等方法,分析了PICK1蛋白N末端区域几个酸性氨基酸残基对PDZ 结构域与膜脂结合的影响,以及钙离子结合N末端酸性区域对PDZ脂结合能力的调节. 结果显示, 带有上游酸性区域的PDZ结构域(NPDZ)的脂质结合能力仅相当PDZ结构 域的15%,相比单独的PDZ结构域与脂质的解离常数Kd(PDZ)为1.58×103 μg·L-1, NPDZ与脂质解离常数Kd(NPDZ)为3.3×104μg·L-1,其中在N末端酸性残基中D8与 D12两个天冬氨酸是影响脂质结合能力减弱的关键残基,若将二者分别突变为丙氨酸 后,NPDZ与脂质的解离常数分别为:Kd (D8/A)=4.42×103μg·L-1;Kd (D12/A) =1.73×103μg·L-1接近于PDZ结构域与脂质结合能力;钙离子会增强NPDZ脂结合能力,当钙离子浓度达到30 μmol/L时,NPDZ的脂结合能力提高2.3倍,但只相当于PDZ的50% 的结合能力.

Abstract

Protein interacting with Cα kinase 1 (PICK1), plays an important role in linking the membrane receptors and PKCα in synapse. We have analyzed the interaction between the PDZ domain and membrane lipid regulated by its adjacent N-terminal acid region, using fluorescence spectra, gene manipulation combined with PLO (protein-lipid overlay assay). The result indicated that N-terminal acid region of PICK1 decreased the interaction between PDZ domain of PICK1 and membrane lipid about 85% degree. D8, D12 of N-terminal acidic regions constitute the key residues for decreasing PDZ binding to lipid. 30 μmol/L Ca2+ could enhance the lipid binding ability of PDZ domain with its adjacent N- terminal acid region about 2.3 times, but that was about 50% degree compared to that of only PDZ domain.

关键词

PDZ结构域 /  N端酸性区域  /   脂质体 /   荧光光谱 / Ca2+

Key words

PDZ domain /   N-terminal acid region /  liposome  /   fluorescence emission spectra  /    / Ca2+

引用本文

导出引用
杨廷, 贾原, 肖虹, 石亚伟. PICK1中N端酸性区域对其PDZ结构域脂质结合能力的调节[J]. 中国生物化学与分子生物学报, 2012, 28(10): 940-945
YANG Ting, JIA Yuan, XIAO Hong, SHI Ya-Wei. Regulation of Liposome Binding of PDZ Domain by Its Adjacent N-terminal Acidic Region[J]. Chinese Journal of Biochemistry and Molecular Biology, 2012, 28(10): 940-945
中图分类号: Q71   

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基金

国家自然科学基金项目(No. 30400065,No. 31170748)和山西省自然科学基金项目资助(No. 2007021033)

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