title: Phosphorylation and function of DGAT1 in skeletal muscle cells.
abstract: Aberrant intramuscular triacylglycerol (TAG) storage in human skeletal muscle is closely related to insulin insensitivity. Excessive lipid storage can induce insulin resistance of skeletal muscle, and under severe conditions, lead to type 2 diabetes. The balance of interconversion between diacylglycerol and TAG greatly influences lipid storage and utilization. Diacylglycerol O-acyltransferase 1 (DGAT1) plays a key role in this process, but its activation and phosphorylation requires further dissection. In this study, 12 putative conserved phosphorylation sites of DGAT1 were identified by examining amino acid conservation of DGAT1 in different species. Another 12 putative phosphorylation sites were also found based on bioinformatics predictions and previous reports. Meanwhile, several phosphorylation sites of DGAT1 were verified by phosphorylation mass spectrometry analysis of purified DGAT1 from mouse myoblast C2C12 cells. Using single point mutations, a regulatory role of 3 putative phosphorylation sites was dissected. Finally, using truncation mutations, a potential domain of DGAT1 that was involved in the regulation of enzymatic activity was revealed. This study provides useful information for further understanding DGAT1 activity regulation.
authors.affiliation: National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, 100101 China ; University of Chinese Academy of Sciences, Beijing, 100049 China.
authors.forename: Jinhai
authors.initials: J
authors.lastname: Yu
authors.affiliation: National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, 100101 China ; Department of Biological Science and Biotechnology, School of Biological Science and Medical Engineering, Beihang University, Beijing, 100191 China.
authors.forename: Yiran
authors.initials: Y
authors.lastname: Li
authors.affiliation: Graduate School of Anhui Medical University, Hefei, 230032 China ; General Hospital of Air Force, Beijing, 100142 China.
authors.forename: Fei
authors.initials: F
authors.lastname: Zou
authors.affiliation: National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, 100101 China ; University of Chinese Academy of Sciences, Beijing, 100049 China.
authors.forename: Shimeng
authors.initials: S
authors.lastname: Xu
authors.affiliation: National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, 100101 China.
authors.forename: Pingsheng
authors.initials: P
authors.lastname: Liu
keywords: DGAT1; Enzymatic activity; Phosphorylation; Single point mutation