Objective: Batten disease （BD）, the juvenile form of neuronal ceroid lipofuscinosis （NCLs）, is pathological characterized by finding lysosomal storage of autofluorescent lipofuscins with unique ultrastructural pro...Objective: Batten disease （BD）, the juvenile form of neuronal ceroid lipofuscinosis （NCLs）, is pathological characterized by finding lysosomal storage of autofluorescent lipofuscins with unique ultrastructural profiles. The gene underlying BD is designated CLN3 and encodes a protein, Battenin, of unknown function that localizes in lysosomes and/or mitochondria. Previously, we hypothesized that Battenin associates with other membrane protein（s） to form a membrane complex. Dysfunction of this complex could result in the pathological changes of BD, and possibly in other NCLs. Two such membranous proteins, the slow and fast Battenin-interactive proteins （BIPs and BIPf） of unknown functions, have been identified. In this study, we have characterized the functional domains of Battenin that interact with both BIP proteins. Methods: Protein-protein interactions with a yeast two-hybrid system were employed. A “deletion assay” was employed to localize the interactive segment（s）. Different lengths of cDNA sequences lacking exon 1-5 were used to express CLN3-encoded proteins lacking N-terminal segments in the yeast two-hybrid system. N-terminal exons of CLN3 were deleted with PCR-cloning strategies.Results: We eliminated the possibility of interacting domains from the exon 7-encoded region because both Battenin and mBattenin interact with the BIP proteins. We have shown that peptide sequences encoded by exons 2 and 4 of CLN3 gene include the functional domains by which Battenin interacts with the BIP proteins. Conclusion: Our studies provide evidence that the N-terminus of Battenin is the functional domain for these protein interactions.展开更多
肺表面活性物质是分布于肺泡内衬的一类磷脂蛋白复合物。而由于磷脂具有两亲性,能与肺表面活性物质中的特异性蛋白相互作用而在肺泡中排列形成磷脂单分子层,从而发挥其降低气液界面表面张力的作用。肺表面活性蛋白B(surfactant protein ...肺表面活性物质是分布于肺泡内衬的一类磷脂蛋白复合物。而由于磷脂具有两亲性,能与肺表面活性物质中的特异性蛋白相互作用而在肺泡中排列形成磷脂单分子层,从而发挥其降低气液界面表面张力的作用。肺表面活性蛋白B(surfactant protein B,SP-B)是肺表面活性蛋白最为重要的特异性蛋白,它的异常常常引起肺部各种疾病,因此SP-B在肺功能维持和修复中起着不可替代的作用。SP-B具有类似于阿米巴穿孔肽超蛋白家族成员的结构,属于鞘脂激活蛋白样蛋白超家族蛋白(SAPLIPs)。通过分析脂质-蛋白相互作用和SP-B的构效关系特点,为基于类似物的药物开发提供理论依据。展开更多
文摘Objective: Batten disease （BD）, the juvenile form of neuronal ceroid lipofuscinosis （NCLs）, is pathological characterized by finding lysosomal storage of autofluorescent lipofuscins with unique ultrastructural profiles. The gene underlying BD is designated CLN3 and encodes a protein, Battenin, of unknown function that localizes in lysosomes and/or mitochondria. Previously, we hypothesized that Battenin associates with other membrane protein（s） to form a membrane complex. Dysfunction of this complex could result in the pathological changes of BD, and possibly in other NCLs. Two such membranous proteins, the slow and fast Battenin-interactive proteins （BIPs and BIPf） of unknown functions, have been identified. In this study, we have characterized the functional domains of Battenin that interact with both BIP proteins. Methods: Protein-protein interactions with a yeast two-hybrid system were employed. A “deletion assay” was employed to localize the interactive segment（s）. Different lengths of cDNA sequences lacking exon 1-5 were used to express CLN3-encoded proteins lacking N-terminal segments in the yeast two-hybrid system. N-terminal exons of CLN3 were deleted with PCR-cloning strategies.Results: We eliminated the possibility of interacting domains from the exon 7-encoded region because both Battenin and mBattenin interact with the BIP proteins. We have shown that peptide sequences encoded by exons 2 and 4 of CLN3 gene include the functional domains by which Battenin interacts with the BIP proteins. Conclusion: Our studies provide evidence that the N-terminus of Battenin is the functional domain for these protein interactions.
文摘肺表面活性物质是分布于肺泡内衬的一类磷脂蛋白复合物。而由于磷脂具有两亲性,能与肺表面活性物质中的特异性蛋白相互作用而在肺泡中排列形成磷脂单分子层,从而发挥其降低气液界面表面张力的作用。肺表面活性蛋白B(surfactant protein B,SP-B)是肺表面活性蛋白最为重要的特异性蛋白,它的异常常常引起肺部各种疾病,因此SP-B在肺功能维持和修复中起着不可替代的作用。SP-B具有类似于阿米巴穿孔肽超蛋白家族成员的结构,属于鞘脂激活蛋白样蛋白超家族蛋白(SAPLIPs)。通过分析脂质-蛋白相互作用和SP-B的构效关系特点,为基于类似物的药物开发提供理论依据。
