iProXProteomicsShixian LinHomo SapiensMus Musculushttp://www.iprox.org/page/project.html?id=IPX0008686000sxlin@zju.edu.cnZhejiang UniversityiProXThe liver proteome undergoes dynamic changes while performing hundreds of essential biological functions. Dysregulation of the liver proteome under alcoholic conditions leads to alcohol-associated liver disease (ALD), a major health challenge worldwide. There is an urgent need for quantitative and liver-specific proteome information in living animals to understand the pathophysiological dynamics of this largest solid organ. Here, we develop a comprehensive approach that specifically identifies the nascent proteome and preferentially enriches membrane proteins in living mouse hepatocytes and is broadly applicable to studies of the liver under various physiological and pathological conditions. In the ethanol-induced liver injury mouse model, the nascent proteome successfully identifies and validates a number of transcription regulators, enzymes, and protective chaperones involved in the molecular regulation of hepatic steatosis, in addition to almost all known regulatory proteins and pathways related to alcohol metabolism. We discover that Phb1/2 is an important transcription coregulator in the process of ethanol metabolism, and one identified fatty acid metabolism enzyme Acsl1/5, whose inhibition protects cells and mice from lipid accumulation, a key symptom of hepatic steatosis.Nascent liver proteome reveals enzymes and transcription regulators under physiological and alcohol exposure conditions.Gu Jiayu J, Lao Lihui L, Hu Linzhen L, Zang Jia J, Liu Chao C, Wan Ruixi R, Tang Ling L, Yuan Ying Y, Chen Yulin Y, Lin Shixian SfalseNascent proteome reveals key regulators in the development of alcoholic liver diseaseAlcoholic liver disease (ALD) poses a significant global health challenge, and the global proteome will change dynamically and dramatically for the response to alcohol stress. However, due to the lack of method for labeling and identification of tissue-specific nascent proteome in living mouse, the underlying molecular mechanism of the pathogenesis remains poorly understood. Here, we developed the stochastic orthogonal recoding of translation induced by AAV-delivered Cre (SORT-AC) strategy to precisely identify the nascent proteome in the hepatocyte of living mice. We applied this strategy to investigate the nascent proteome and underlying molecular mechanism in ALD mouse model. We identified 187 newly synthesized proteins involved in protein and metabolite processing in liver of ALD model mice. We found that upregulated Phb1/2 is responsible for the transcription of ethanol metabolism related genes and the downstream substrate, Hsp family proteins were responsible to protect from misfolded protein aggregation. Another downstream substrate of Phb1/2, Acsl1/5 participated in the lipid accumulation. Furthermore, the inhibition of Acsl1/5 protected cultured cells and mice from lipid accumulation by alcohol exposure, which might be a brand-new therapeutic target to antagonist the hepatocyte damage of ALD. More broadly, these results pave the new way for the nascent proteome investigation in living animal models in a tissue-specific manner.Tue May 20 00:00:00 GMT+01:00 2025PXD05172510090960640858584