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Systematic proteomics and biochemical approaches to identifies the function of UBXN10 in p97‐associated process
Author(s) -
li xiaolong,
li shan,
Chou TsuiFen
Publication year - 2016
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.30.1_supplement.610.2
Subject(s) - aaa proteins , endoplasmic reticulum associated protein degradation , endoplasmic reticulum , microbiology and biotechnology , ubiquitin , subcellular localization , biology , mutant , immunoprecipitation , ultrabithorax , function (biology) , atpase , biochemistry , chemistry , unfolded protein response , enzyme , gene , homeotic gene , cytoplasm
The VCP AAA‐ATPase (also known as p97), an evolutionarily conserved ATP‐driven ‘segregase’, is a central regulator of protein quality control and ubiquitin‐mediated signalling. VCP functions in diverse processes such as endoplasmic reticulum (ER)‐associated degradation (ERAD), macro and selective autophagy and DNA damage responses(DDRs). These functions are dependent on association with cofactors that are specific to the different biological pathways p97 participates in. The UBX‐protein family (ubiquitin regulatory X) is an evolutionary conserved protein family and determine the specificity of p97/VCP/Cdc48p function by binding as its adaptors. constitutes the largest known group of p97 cofactors. However, few general or specific function of them has been revealed. Especially UBXD3, During the course of understanding not only their function but also specified function of p97, we investigated how the UBXD3 and the protein bind with UBXD3 worked in this study. The steady‐state kinetic study demonstrates that UBXD3 increases the catalytic efficiency (Kcat/Km) of both WT and the D2‐active mutant, but does not affect the D1‐active mutant. Thus, UBXD3 mainly regulates the D2 ATPase activity of p97. We used the co‐immunoprecipitation (1P) and LC‐MS/MS quantitation approaches to analyze interacting proteins of p97 and UBXD3. Analysis of UBXD3—Flag IP from transient transfection with 293T and U20S cells demonstrated that UBXD3 could bind with MOB1B and STK3. Indicate the function of UBXD3 is related to the Hippo signaling pathway.