title: Human intersectin 2 (ITSN2) binds to Eps8 protein and enhances its degradation.
abstract: Participates in actin remodeling through Rac and receptor endocytosis via Rab5. Here, we used yeast two-hybrid system with Eps8 as bait to screen a human brain cDNA library. ITSN2 was identified as the novel binding factor of Eps8. The interaction between ITSN2 and Eps8 was demonstrated by the in vivo co-immunoprecipitation and colocalization assays and the in vitro GST pull-down assays. Furthermore, we mapped the interaction domains to the region between amino acids 260-306 of Eps8 and the coiled-coil domain of ITSN2. In addition, protein stability assays and immunofluorescence analysis showed ITSN2 overexpression induced the degradation of Eps8 proteins, which was markedly alleviated with the lysosome inhibitor NH4Cl treatment. Taken together, our results suggested ITSN2 interacts with Eps8 and stimulates the degradation of Eps8 proteins.
authors.affiliation: Key Laboratory of Protein Chemistry and Development Biology of State Education Ministry of China, College of Life Science, Hunan Normal University, Changsha, China.
authors.forename: Xiaofeng
authors.initials: X
authors.lastname: Ding
authors.forename: Zijian
authors.initials: Z
authors.lastname: Yang
authors.forename: Fangliang
authors.initials: F
authors.lastname: Zhou
authors.forename: Xiang
authors.initials: X
authors.lastname: Hu
authors.forename: Chang
authors.initials: C
authors.lastname: Zhou
authors.forename: Chang
authors.initials: C
authors.lastname: Luo
authors.forename: Zhicheng
authors.initials: Z
authors.lastname: He
authors.forename: Qian
authors.initials: Q
authors.lastname: Liu
authors.forename: Hong
authors.initials: H
authors.lastname: Li
authors.forename: Feng
authors.initials: F
authors.lastname: Yan
authors.forename: Fangmei
authors.initials: F
authors.lastname: Wang
authors.forename: Shuanglin
authors.initials: S
authors.lastname: Xiang
authors.forename: Jian
authors.initials: J
authors.lastname: Zhang
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism*; D033942:Adaptor Proteins, Vesicular Transport / Q000378:metabolism*; D001665:Binding Sites; D006801:Humans; D011485:Protein Binding; D059748:Proteolysis
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D033942:Adaptor Proteins, Vesicular Transport; C489989:EPS8 protein, human; C509776:ITSN2 protein, human