Kang, Joo Ae
(Metabolic Regulation Research Center, KRIBB)
,
Kang, Hyun Sup
(Metabolic Regulation Research Center, KRIBB)
,
Bae, Kwang-Hee
(Metabolic Regulation Research Center, KRIBB)
,
Lee, Sang Chul
(Metabolic Regulation Research Center, KRIBB)
,
Oh, Kyoung-Jin
(Metabolic Regulation Research Center, KRIBB)
,
Kim, Won Kon
(Metabolic Regulation Research Center, KRIBB)
Despite the importance of brown adipocytes as a therapeutic target for the prevention and treatment of obesity, the molecular mechanism underlying brown adipocyte differentiation is not fully understood. In particular, the role of post-translational modifications in brown adipocyte differentiation h...
Despite the importance of brown adipocytes as a therapeutic target for the prevention and treatment of obesity, the molecular mechanism underlying brown adipocyte differentiation is not fully understood. In particular, the role of post-translational modifications in brown adipocyte differentiation has not been extensively studied. Histidine phosphorylation is increasingly recognized an important process for protein post-translational modifications. In this study, we show that histidine phosphorylation patterns change during brown adipocyte differentiation. In addition, the expression level of protein histidine phosphatase 1 (PHPT1), a major mammalian phosphohistidine phosphatase, is reduced rapidly at the early phase of differentiation and recovers at the later phase. During white adipocyte differentiation of 3T3-L1 preadipocytes, however, the expression level of PHPT1 do not significantly change. Knockdown of PHPT1 promotes brown adipocyte differentiation, whereas ectopic expression of PHPT1 suppresses brown adipocyte differentiation. These results collectively suggest that histidine phosphorylation is closely linked to brown adipocyte differentiation and could be a therapeutic target for obesity and related metabolic diseases.
Despite the importance of brown adipocytes as a therapeutic target for the prevention and treatment of obesity, the molecular mechanism underlying brown adipocyte differentiation is not fully understood. In particular, the role of post-translational modifications in brown adipocyte differentiation has not been extensively studied. Histidine phosphorylation is increasingly recognized an important process for protein post-translational modifications. In this study, we show that histidine phosphorylation patterns change during brown adipocyte differentiation. In addition, the expression level of protein histidine phosphatase 1 (PHPT1), a major mammalian phosphohistidine phosphatase, is reduced rapidly at the early phase of differentiation and recovers at the later phase. During white adipocyte differentiation of 3T3-L1 preadipocytes, however, the expression level of PHPT1 do not significantly change. Knockdown of PHPT1 promotes brown adipocyte differentiation, whereas ectopic expression of PHPT1 suppresses brown adipocyte differentiation. These results collectively suggest that histidine phosphorylation is closely linked to brown adipocyte differentiation and could be a therapeutic target for obesity and related metabolic diseases.
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가설 설정
reported a total of 786 potential phosphohistidine substrates by proteomics analysis approaches using sequence-independent monoclonal pHis antibody [8]. In light of the fact that the number of potential substrates were much larger than expected, we hypothesized that histidine phosphorylation would be more critically associated with biological processes than previously thought.
As such, the effects of histidine phosphorylation in brown adipocytes function or differentiation have not been identified yet. In this study, we investigated whether histidine phosphorylation affects brown adipocyte differentiation. We also analyzed the involvement of PHPT1, which regulates histidine phosphorylation during brown adipocyte differentiation.
제안 방법
To further clarify the roles of PHPT1 in brown adipogenesis, we analyzed the effect of the ectopic expression of PHPT1 during brown adipocyte differentiation.
3B and 3E). To investigate the function of PHPT1 in brown adipocyte differentiation, PHPT1 knockdown cells were induced to differentiate for six days. PHPT1 knockdown promoted differentiation leading to increased lipid droplet accumulation in comparison to that of control cells (Fig.
데이터처리
Statistical analysis was performed using an independent Student’s t-test (***p < 0.0005).
Statistical analysis was performed using an independent Student’s t-test.
성능/효과
Second, PHPT1 is suppressed at an early phase and it recovers at a later stage of differentiation, implying the involvement of PHPT1 in brown adipocyte differentiation through the modulation of the protein histidine phosphorylation. Third, both the knockdown and ectopic expression of PHPT1 had significant effects on brown adipocyte differentiation. Although the current studies were limited to PHPT1 among the three phosphohistidine phosphatases and the direct target substrate(s) of PHPT1 remain to be identified, our results reveal a significant role for protein histidine phosphorylation and PHPT1 during brown adipocyte differentiation.
후속연구
However, there are only a few reports regarding the specific substrates of each histidine kinase and phosphatase. Thus, more detailed studies are necessary to understand the cellular mechanisms of histidine phosphorylation.
4F), implying that PHPT1 might regulate brown adipocyte differentiation by modulating histidine phosphorylation of ACLY. However, further studies are needed to elucidate the detailed mechanism between histidine phosphorylation of ACLY and brown adipogenic differentiation. Previously, Seeger et al.
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