PICK1中N端酸性区域对其PDZ结构域脂质结合能力的调节

作者: 时间:2025-01-18 点击量:

1)教育部化学生物学与分子工程重点实验室,山西大学生物技术研究所,太原030006;2)山西医学科学院生物治疗科中心实验室,太原030006;3)山西医科大学第一医院病理科,太原030001 1)Institute of Biotechnology, Key Laboratory of Chemical Biology and Molecular Engineering of Education Ministry, Shanxi University, Taiyuan030006, China;2)Department of Biotherapy, Shanxi Academy of Medical Sciences, Taiyuan030006, China;3 )Department of Pathology, First Affiliated Hospital, Shanxi Medical University, Taiyuan030001, China 摘要蛋白激酶C 相互作用蛋白1(protein interacting with C kinase 1, PICK1)是衔接膜上受体和蛋白激酶C 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 g L-1, NPDZ与脂质解离常数Kd(NPDZ)为3.3 104 g 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)=4.42 103 g L-1;Kd (D12/A) =1.73 103 g 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 mol/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 C kinase 1 (PICK1), plays an important role in linking the membrane receptors and PKC 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 mol/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. 基金资助: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) 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. Chinese Journal of Biochemistry and Molecular Biol, 2012, 28(10): 940-945. Hanley JG., Henley JM. PICK1 is a calcium-sensor for NMDA-induced AMPA receptor trafficking[J]. EMBO J,2005,24(18):3266-3278 Beattie EC,Carroll RC,Yu X,et al. Regulation of AMPA receptor endocytosis by a signaling mechanism shared with LTD[J].Nat Neurosci,2000,3(12):1291-1300 Citri A, Bhattacharyya S, Ma C,et al. Calcium binding to PICK1 is essential for the intracellular retention of AMPA receptors underlying long-term depression[J]. J Neurosci,2010,30(49):16437-16452 Liao D, Scannevin RH, Huganir R. Activation of silent synapses by rapid activity-dependent synaptic recruitment of AMPA receptors[J].J Neurosci,2001,21(16): 6008-17 Lin RC, Scheller RH. Mechanisms of synaptic vesicle exocytosis[J]. Annu Rev Cell Dev Biol,2000,16: 19-49 Mochida S. Protein-protein interactions in neurotransmitter release[J].Neurosci Res,2000,36(3):175-182 Bolia A, Gerek ZN, Keskin O, et al. The binding affinities of proteins interacting with the PDZ domain of PICK1[J].Proteins,2012,80(5):1393-1408 Ammendrup-Johnsen l, Thorsen TS, Gether U, et al. Serine 77 in the PDZ domain of PICK1 is a protein kinase C phosphorylation site regulated by lipid membrane binding[J].Biochemistry,2012,51(2):586-596 肖虹,袁静明,石亚伟. 蛋白激酶C 相互作用蛋白的结构与功能[J].细胞生物学杂志(Xiao Hong, Yuan Jing-Ming , Shi Ya-Wei. The structure and function of protein interacting with C kinase [J].Chin J Cell Biol),2006, 28:137-140 Shao X,Zhu L,Wang Y,Lu Y,et al. Threonine 82 at the PDZ domain of PICK1 is critical for AMPA receptor interaction and localization[J].Neurochem Int,2010,56(8):962-970 Xia J,Zhang X, Staudinger J, et al. Clustering of AMPA receptors by the synaptic PDZ domain-containing protein PICK1[J].Neuron,1999,22(1):179-187 Hanley JG. NSF binds calcium to regulate its interaction with AMPA receptor subunit GluR2[J].J Neurochem,2007,101(6):1644-1650 Citri A, Bhattacharyya S, Ma C, et al. Calcium binding to PICK1 is essential for the intracellular retention of AMPA receptors underlying long-term depression[J].J Neurosci,2010,30(49):16437-52 贾原,黄玉民,刘军林,等. PICK1蛋白C端酸性区域对其功能的调节[J].中国生物化学与分子生物学报( Jia Yuan, Huang Yu-Min, Liu Jun-Lin, et al. C-terminal acidic region regulates PICK1 function[J]. Chin J Biochem Mol Biol),2010,26(3):224-230 Pan L,Wu H,Shen C, et al. Clustering and synaptic targeting of PICK1 requires direct interaction between the PDZ domain and lipid membranes[J].EMBO J,2007,26(21):4576-4587 Shi Y, Yu J, Jia Y, et al. Redox-regulated lipid membrane binding of the PICK1 PDZ domain[J]. Biochemistry,2010,49(21):4432-4439 Xiao H, Shi Y, Yuan J, et al. Over-expression, rapid preparation and some properties of C-terminal BARc region in PICK1[J].Int J Mol Sci,2009,10(1):28-36 Bramkamp M , Altendorf K. Functional modules of KdpB,the catalytic subunit of the Kdp-ATPase from Escherichia coli[J].Biochemistry,2004,43(38):12289-12296 石亚伟,张蕾,肖红,等.重组PICK1蛋白的多聚形式及其与Ca2+和Mg2+的结合[J].中国生物化学与分子生物学报(Shi Ya-Wei,Zhang Lei,Xiao Hong,et al.Oligomeric properties of recombinant PICK1 and its binding capacity with Ca2 + and Mg2 + [J]. Chin J Biochem Mol Biol),2007,23(12):1012-1018 Shi Y, Zhang L, Yuan J, et al. Zinc binding site in PICK1 is dominantly located at the CPC motif of its PDZ domain[J].J Neurochem, 2008,106(3):1027-1034 Lu W, Ziff EB. PICK1 interacts with ABP/GRIP to regulate AMPA receptor trafficking[J]. Neuron, 2005,47(3)407-421 王童 隋立乾 康翠洁. 多功能的胞内衔接蛋白syntenin[J]. 中国生物化学与分子生物学报, 2013, 29(12): 1097-1105. 李华, 林葵, 吴更. PDZ结构域的结构特点及其识别特异性配体的机制[J]. 中国生物化学与分子生物学报, 2011, 27(12): 1106-1111. 宋婀莉,高友鹤. PICK1:一个功能多样的药靶蛋白[J]. 中国生物化学与分子生物学报, 2007, 23(09): 697-705. 田瑞,李英娜,宋婀莉,高诗娟,黄海明,张玲,马素参,高友鹤. 通过筛选随机多肽文库研究ZO-1中PDZ1结构域的配体结合特点[J]. 中国生物化学与分子生物学报, 2006, 22(06): 484-490.

>>> 更多资讯详情请访问蚂蚁淘商城

Molecular Dimensions商品列表
图片/货号 产品名/品牌 价格/货期 操作