Deubiquitinase OTUB1 is involved in the cellular response to copper deficiency

• Published in: Ji chu yi xue yu lin chuang = Jichu yixue yu linchuang = Basic medical sciences and clinics Vol. 42; no. 7; pp. 1020 - 1025
• Main Author: ZHANG Yu-fei, ZHU Ji-yan, QU Si-yao, ZHANG Zhu-qin, LIU De-pei
• Format: Journal Article
• Language: Chinese
• Published: Institute of Basic Medical Sciences and Peking Union Medical College Hospital, Chinese Academy of Medical Sciences / Peking Union Medical College 01-07-2022
• Subjects: copper|cardiac hypertrophy|otub1
• ISSN: 1001-6325
• DOI: 10.16352/j.issn.1001-6325.2022.07.1020

Objective By establishing a copper deficiency cell model to explore whether there are copper-dependent enzymes involved in cardiac hypertrophy caused by copper deficiency. Methods The Ctr1 knockout H9c2 cell strain was established by CRISPR/Cas9 technology; The proteins in wildtype (WT), Ctr1-/-, and copper chelator TTM-treated H9c2 cells were collected, and the protein was screened by copper affinity resin pull down experiment combined with mass spectrometry analysis to find out an enzyme which could interact with copper; The downstream signaling pathway of this enzyme in copper-deficient cells was detected by Western blot. Results The H9c2 cell strain with knockout of Ctr1 was successfully established, and the copper level in this cell strain was significantly reduced (P＜0.05). Mass spectrometry analysis screened out the deubiquitinase OTUB1 that could interact with copper; The level of ubiquitination in copper-deficient H9c2 cells was increased; Under the condition of copper deficiency, the level of OTUB1