Ma, Ruiying Pang, Kaifang Kang, Hyojin Zhang, Yinhua Bang, Geul Park, Sangwoo Hwang, Eunha Ryu, Jae Ryun Kwon, Yujin Kang, Hyae Rim Jin, Chunmei Kim, Yoonhee Kim, Su Yeon Kwon, Seok-Kyu Kim, Doyoun Sun, Woong Kim, Jin Young Han, Kihoon 2024-01-19T11:34:04Z 2024-01-19T11:34:04Z 2022-06-17 2022-07 0022-3042 https://pubs.kist.re.kr/handle/201004/114913 The two members of the cytoplasmic FMR1-interacting protein family, CYFIP1 and CYFIP2, are evolutionarily conserved multifunctional proteins whose defects are associated with distinct types of brain disorders. Even with high sequence homology between CYFIP1 and CYFIP2, several lines of evidence indicate their different functions in the brain; however, the underlying mechanisms remain largely unknown. Here, we performed reciprocal immunoprecipitation experiments using CYFIP1-2 x Myc and CYFIP2-3 x Flag knock-in mice and found that CYFIP1 and CYFIP2 are not significantly co-immunoprecipitated with each other in the knock-in brains compared with negative control wild-type (WT) brains. Moreover, CYFIP1 and CYFIP2 showed different size distributions by size-exclusion chromatography of WT mouse brains. Specifically, mass spectrometry-based analysis of CYFIP1-2 x Myc knock-in brains identified 131 proteins in the CYFIP1 interactome. Comparison of the CYFIP1 interactome with the previously identified brain region-and age-matched CYFIP2 interactome, consisting of 140 proteins, revealed only eight common proteins. Investigations using single-cell RNA-sequencing databases suggested non-neuronal cell-and neuron-enriched expression of Cyfip1 and Cyfip2, respectively. At the protein level, CYFIP1 was detected in both neurons and astrocytes, while CYFIP2 was detected only in neurons, suggesting the predominant expression of CYFIP1 in astrocytes. Bioinformatic characterization of the CYFIP1 interactome, and co-expression analysis of Cyfip1 with astrocytic genes, commonly linked CYFIP1 with focal adhesion proteins. Immunocytochemical analysis and proximity ligation assay suggested partial co-localization of CYFIP1 and focal adhesion proteins in cultured astrocytes. Together, these results suggest a CYFIP1-specific association with astrocytic focal adhesion, which may contribute to the different brain functions and dysfunctions of CYFIP1 and CYFIP2. English Blackwell Publishing Inc. Protein interactome and cell-type expression analyses reveal that cytoplasmic FMR1-interacting protein 1 (CYFIP1), but not CYFIP2, associates with astrocytic focal adhesion Article 10.1111/jnc.15622 1 Journal of Neurochemistry, v.162, no.2, pp.190 - 206 Journal of Neurochemistry 162 2 190 206 N scie scopus 000802818300001 Biochemistry & Molecular Biology Neurosciences Biochemistry & Molecular Biology Neurosciences & Neurology Article WAVE FMRP BEHAVIORS NEURONS astrocyte co-expression CYFIP1 CYFIP2 focal adhesion interactome