• 한국자원식물학회지
• /
• 제36권4호
• /
• pp.275-281
• /
• 2023

본 연구에서 우리는 작약 뿌리 추출물이 TLR4/PI3K/Nrf2 신호전달을 통해 p62/SQSTM1을 증가시켜 대식세포에서 자가포식을 유도한다는 것을 확인하였다. 대식세포의 자가포식 유도는 선천성과 적응성 면역 반응 간의 연결을 강화해 백신 보조제 개발에 있어서 중요한 표적으로 사용되기 때문에, 작약 뿌리 추출물은 백신개발에 필수적인 백신보조제로의 활용이 가능할 것으로 생각된다.

• Biodesign
• /
• 제6권4호
• /
• pp.100-102
• /
• 2018

Autophagy is a degradation pathway that targets many cellular components and plays a particularly important role in protein degradation and recycling. This process is very complex and several proteins participate in this process. One of them, P62/SQSTM1, is related to the N-end rule and induces protein degradation through autophagy. The P62/SQSTM1 makes a huge oligomer, and this oligomerization is known to play an important role in its mechanism. This oligomerization takes two steps. First, the PB1 domain of P62/SQSTM1 makes the base oligomer, and then, when the ligand binds to the ZZ domain of P62/SQSTM1, it induces a higher oligomer by the disulfide bond of the two cysteines. To understand the oligomerization mechanism of P62/SQSTM1, we need to know the dimerization of the PB1 domain. In this study, crystals of PB1 dimer were made and the crystals were diffracted by X-ray to collect usable data up to 3.2A. We are analyzing the structure using the molecular replacement (MR) method.

• Asian Pacific Journal of Cancer Prevention
• /
• 제14권7호
• /
• pp.4095-4099
• /
• 2013

Oxidative stress induces apoptosis in many cellular systems including glioblastoma cells, with caspase-8 activation was regarded as a major contribution to $H_2O_2$-induced cell death. This study focused on the role of the autophagic protein p62 in $H_2O_2$-induced apoptosis in U87MG cells. Oxidative stress was applied with $H_2O_2$, and cell apoptosis and viability were measured with use of caspase inhibitors or autophagic mediators or siRNA p62, GFP-p62 and GFP-p62-UBA (del) transfection. We found that $H_2O_2$-induced U87MG cell death was correlated with caspase-8. To understand the role of p62 in MG132-induced cell death, the levels of p62/SQSTM1 or autophagy in U87MG cells were modulated with biochemical or genetic methods. The results showed that the over-expression of wild type p62/SQSTM1 significantly reduced $H_2O_2$ induced cell death, but knockdown of p62 aggravated the process. In addition, inhibition of autophagy promoted p62 and active caspase-8 increasing $H_2O_2$-induced apoptosis while induction of autophagy manifested the opposite effect. We further demonstrated that the function of p62/SQSTM1 required its C-terminus UBA domain to attenuate $H_2O_2$ cytotoxity by inhibition of caspase-8 activity. Our results indicated that p62/SQSTM1 was a potential contributor to mediate caspase-8 activation by autophagy in oxidative stress process.

• Molecules and Cells
• /
• 제42권10호
• /
• pp.729-738
• /
• 2019

Autophagy is an important process for protein recycling. Oligomerization of p62/SQSTM1 is an essential step in this process and is achieved in two steps. Phox and Bem1p (PB1) domains can oligomerize through both basic and acidic surfaces in each molecule. The ZZ-type zinc finger (ZZ) domain binds to target proteins and promotes higher-oligomerization of p62. This mechanism is an important step in routing target proteins to the autophagosome. Here, we determined the crystal structure of the PB1 homo-dimer and modeled the p62 PB1 oligomers. These oligomer models were represented by a cylindrical helix and were compared with the previously determined electron microscopic map of a PB1 oligomer. To accurately compare, we mathematically calculated the lead length and radius of the helical oligomers. Our PB1 oligomer model fits the electron microscopy map and is both bendable and stretchable as a flexible helical filament.

• 대한의생명과학회지
• /
• 제27권3호
• /
• pp.182-186
• /
• 2021

Parkinson disease (PD) is becoming one of the most neurodegenerative disorder worldwide. The deposited aggregates have been connected in the pathophysiology of PD, which are degraded either by ubiquitin-proteasomal system (UPS) or autophagy-lysosomal pathway (ALP). Leucin-rich repeat kinase 2 (LRRK2), one of the neurodegenerative proteins of PD is also stringently controlled by both UPS and ALP degradation as well. However, the polyubiquitination pattern of LRRK2 aggregates is largely unknown. Here, we found that K63-linked polyubiquitinations of G2019S mutant, most familial variant for PD, is highly enhanced compared to those of wild type LRRK2 (WT). In addition, in the presence of overexpressed p62/SQSTM-1, ubiquitination of LRRK2 WT or D1994A was reduced, whereas G2019S mutant was not diminished significantly. Therefore, we propose that degradation of G2019S via UPS is more involved with K63-linked ubiquitination than K48-linked ubiquitination, and overexpressed p62/SQSTM-1 does not enhance degradative effect on G2019S variant.

• 한국자원식물학회지
• /
• 제36권3호
• /
• pp.191-197
• /
• 2023

Autophagy contributes to enhancing the immune system (innate and adaptive immune system) against foreign pathogens. Autophagy of macrophages is used as a major indicator for developing vaccine adjuvants to increase the adaptive immune response. In this study, HSL increased p62/SQSTM1 expression. Inhibition of TLR4, p38, JNK, and NF-κB blocked HSL-mediated increase of p62/SQSTM1. HSL activated p38, JNK, and NF-κB signaling, but HSL-mediated activation of p38, JNK, and NF-κB signaling was reversed by TLR4 inhibition. In addition, HSL increased Nrf2 expression, but HSL-mediated Nrf2 expression did not occur in the inhibition of TLR4, p38, JNK, and NF-κB. Taken together, it is believed that HSL-mediated autophagy may be dependent on activating Nrf2 expression via TLR4-dependent activation of p38, JNK, and NF-κB in macrophages.

• 한국자원식물학회:학술대회논문집
• /
• 한국자원식물학회 2023년도 임시총회 및 춘계학술대회
• /
• pp.45-45
• /
• 2023

Autophagy contributes to enhancing the immune system (innate and adaptive immune system) against foreign pathogens. Autophagy of macrophages is used as a major indicator for developing vaccine adjuvants to increase the adaptive immune response. In this study, RAL increased the production of immunostimulatory mediators and phagocytotic activity in RAW264.7 cells. RAL increased p62/SQSTM1 expression. Inhibition of TLR4, JNK, and PI3K/AKT blocked RAL-mediated increase of p62/SQSTM1. RAL activated JNK and PI3K/AKT signaling. RAL-mediated activation of JNK and PI3K/AKT signaling was reversed by TLR4 inhibition. Taken together, it is believed that RAL-mediated autophagy may be dependent on activating via TLR4-dependent activation of JNK and PI3K/AKT signaling in macrophages.

• Journal of Microbiology and Biotechnology
• /
• 제33권7호
• /
• pp.934-940
• /
• 2023

Syneilesis palmata (SP) is a traditional medicinal plant. SP has been reported to have anti-inflammatory, anticancer, and anti-human immunodeficiency virus (HIV) activities. However, there is currently no research available on the immunostimulatory activity of SP. Therefore, in this study, we report that S. palmata leaves (SPL) activate macrophages. Increased secretion of both immunostimulatory mediators and phagocytic activity was observed in SPL-treated RAW264.7 cells. However, this effect was reversed by the inhibition of TLR2/4. In addition, inhibition of p38 decreased the secretion of immunostimulatory mediators induced by SPL, and inhibition of TLR2/4 decreased the phosphorylation of p38 induced by SPL. SPL augmented p62/SQSTM1 and LC3-II expression. The increase in protein levels of p62/SQSTM1 and LC3-II induced by SPL was decreased by the inhibition of TLR2/4. The results obtained from this study suggest that SPL activates macrophages via TLR2/4-dependent p38 activation and induces autophagy in macrophages via TLR2/4 stimulation.

• 한국자원식물학회:학술대회논문집
• /
• 한국자원식물학회 2023년도 임시총회 및 춘계학술대회
• /
• pp.44-44
• /
• 2023

Syneilesis palmata (SP) has been used as a traditional medicinal plant and vegetable. SP was reported to exert pharmacological activities such as anti-inflammation, anti-cancer, and anti-HIV. However, there are no studies on the immunostimulatory activity of SP. Thus, in this study, we report that S. palmata leaves (SPL) induce the activation of macrophages. An increase in both secretions of immunostimulatory mediators and phagocytotic activity was observed in SPL-treated RAW264.7 cells. However, this was reversed by inhibition of TLR2/4. In addition, the p38 inhibition reduced the SPL-mediated secretion of immunostimulatory mediators, and the SPL-mediated p38 activation was blocked by the TLR2/4 inhibition. SPL augmented both p62/SQSTM1 and LC3-II. TLR2/4 inhibition blocked the SPL-mediated increase of p62/SQSTM1 and LC3-II. These findings indicate that SPL may activate macrophages through TLR2/4-dependent p38 activation and activate autophagy through TLR2/4 stimulation.

• 한국자원식물학회:학술대회논문집
• /
• 한국자원식물학회 2022년도 추계학술대회
• /
• pp.93-93
• /
• 2022

Hovenia dulcis, one of the traditional medicinal plants, is currently being used as a functional ingredient for the development of health functional foods that protects the liver from alcohol damage in Korea. A variety of pharmacological effects of Hovenia dulcis have been reported so far, but studies on immune-enhancing activity are insufficient. Thus, in this study, we report that Hovenia dulcis branches (HDB) induce the activation of macrophages. HDB increased the production of immunostimulatory factors and phagocytosis in RAW264.7 cells. TLR4 inhibition blocked HDB-mediated production of immunostimulatory factors. In addition, the JNK inhibition reduced the HDB-mediated production of immunostimulatory factors, and the HDB-mediated JNK activation was blocked by the TLR4 inhibition. HDB increased the level of LC3-II and p62/SQSTM1. TLR4 inhibition blocked HDB-mediated increase in the level of LC3-II and p62/SQSTM1. These findings indicate that HDB may induce TLR4/JNK-dependent macrophage activation and TLR4-dependent macrophage autophagy.