• The Korean Journal of Physiology and Pharmacology
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• 제10권3호
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• pp.155-159
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• 2006

Among the Shc proteins, p66shc is known to be related to oxidative stress responses and regulation of the production of reactive oxygen species (ROS). The present study was undertaken to investigate the role of p66shc on endothelial nitric oxide synthase (eNOS) activity in the mouse embryonic fibroblasts (MEFs). When wild type (WT) or p66shc (-/-) MEFs were transfected with full length of eNOS cDNA, the expression and activity of eNOS protein were higher in the p66shc (-/-) MEFs. These phenomena were reversed by reconstitution of p66shc cDNA transfection in the p66shc (-/-) MEFs. The basal superoxide production in the p66shc (-/-) MEFs was not significantly different from that of WT of MEFs. However, superoxide production induced by NADPH in the p66shc (-/-) MEF was lesser than that in WT MEFs. When compared with WT MEFs, cell lysate of p66shc (-/-) MEFs showed significantly increased H-ras activity without change of endogenous H-ras expression. Our findings suggest the pivotal role of p66shc adaptor protein played in inhibition of endothelial nitric oxide production via modulation of the expression and/or activity of eNOS protein.

• Animal Bioscience
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• 제36권7호
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• pp.1022-1033
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• 2023

Objective: p66Shc, a 66 kDa protein isoform encoded by the proto-oncogene SHC, is an essential intracellular redox homeostasis regulatory enzyme that is involved in the regulation of cellular oxidative stress, apoptosis induction and the occurrence of multiple age-related diseases. This study investigated the expression profile and functional characteristics of p66Shc during preimplantation embryo development in sheep. Methods: The expression pattern of p66Shc during preimplantation embryo development in sheep at the mRNA and protein levels were studied by quantitative real-time polymerase chain reaction (RT-qPCR) and immunofluorescence staining. The effect of p66Shc knockdown on the developmental potential were evaluated by cleavage rate, morula rate and blastocyst rate. The effect of p66Shc deficiency on reactive oxygen species (ROS) production, DNA oxidative damage and the expression of antioxidant enzymes (e.g., catalase and manganese superoxide dismutase [MnSOD]) were also investigated by immunofluorescence staining. Results: Our results showed that p66Shc mRNA and protein were expressed in all stages of sheep early embryos and that p66Shc mRNA was significantly downregulated in the 4-to 8-cell stage (p<0.05) and significantly upregulated in the morula and blastocyst stages after embryonic genome activation (EGA) (p<0.05). Immunofluorescence staining showed that the p66Shc protein was mainly located in the peripheral region of the blastomere cytoplasm at different stages of preimplantation embryonic development. Notably, serine (Ser36)-phosphorylated p66Shc localized only in the cytoplasm during the 2- to 8-cell stage prior to EGA, while phosphorylated (Ser36) p66Shc localized not only in the cytoplasm but also predominantly in the nucleus after EGA. RNAi-mediated silencing of p66Shc via microinjection of p66Shc siRNA into sheep zygotes resulted in significant decreases in p66Shc mRNA and protein levels (p<0.05). Knockdown of p66Shc resulted in significant declines in the levels of intracellular ROS (p<0.05) and the DNA damage marker 8-hydroxy2'-deoxyguanosine (p<0.05), markedly increased MnSOD levels (p<0.05) and resulted in a tendency to develop to the morula stage. Conclusion: These results indicate that p66Shc is involved in the metabolic regulation of ROS production and DNA oxidative damage during sheep early embryonic development.

• The Korean Journal of Physiology and Pharmacology
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• 제16권3호
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• pp.199-204
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• 2012

We evaluated the role of Tat-mediated p66shc transduction on the activation of endothelial nitric oxide synthase in cultured mouse endothelial cells. To construct the Tat-p66shc fusion protein, human full length p66shc cDNA was fused with the Tat-protein transduction domain. Transduction of TAT-p66shc showed a concentration- and time-dependent manner in endothelial cells. Tat-mediated p66shc transduction showed increased hydrogen peroxide and superoxide production, compared with Tat-p66shc (S/A), serine 36 residue mutant of p66shc. Tat-mediated p66shc transduction decreased endothelial nitric oxide synthase phosphorylation in endothelial cells. Furthermore, Tat-mediated p66shc transduction augmented TNF-${\alpha}$-induced p38 MAPK phosphorylation in endothelial cells. These results suggest that Tat-mediated p66shc transduction efficiently inhibited endothelial nitric oxide synthase phosphorylation in endothelial cells.

• IMMUNE NETWORK
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• 제6권2호
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• pp.67-75
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• 2006

Background: Cytotoxic function of killer cells is inhibited by specific recognition of class I MHC molecules on target cells by inhibitory killer Ig-like receptors (KIR) expressed on NK cells and some cytotoxic T cells. The inhibitory effect of KIR is accomplished by recruitment of SH2-containing protein tyrosine phosphatase (SHP) to the phosphotyrosine residues in the cytoplasmic tail. Methods: By in vitro coprecipitation experiments and transfection analysis, we investigated the association of KIR with an adaptor protein Shc in Jurkat T cells. Results: The cytoplasmic tail of KIR appeared to associate with an adaptor protein Shc in Jurkat T celilysates. Similar in vitro experiments showed that phosphorylated KIR cytoplasmic tail bound SHP-1 and Shc in Jurkat T cell lysates. The association of KIR with Shc was further confirmed by transfection analysis in 293T cells. Interestingly, however, Shc appeared to be replaced by SHP-2 upon engagement of KIR in 293T cells. Conclusion: Our data indicate that KIR associate with an adaptor protein Shc in Jurkat T cells, and suggest that KIR might have an additional role which is mediated by this adaptor protein.

• 미생물학회지
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• 제55권2호
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• pp.123-130
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• 2019

Chelatase는 tetrapyrrole에 2가 금속을 삽입하는 데 관여하는 효소로서 cobalamin, siroheme, heme, chlorophyll과 같은 금속-tetrapyrrole의 생합성에 필수적인 역할을 담당한다. SirB는 sirohydrochlorin(SHC) tetrapyrrole의 중앙부에 코발트나철을 삽입하여 코발트-SHC 또는 철-SHC를 형성하는 SHC chelatase이다. SirB의 금속 결합 기전 및 SHC 인식 기전을 구조적으로 이해하기 위해 Bacillus subtilis subsp. spizizenii에서 유래한 SirB(bssSirB)의 코발트 복합체 구조를 규명하였다. bssSirB는 N-말단 도메인(NTD)과 C-말단 도메인(CTD)으로 구성된 ${\alpha}/{\beta}$ 단량체 구조를 형성한다. bssSirB는 NTD와 CTD 사이에 서열 보존성이 높은 공동을 지니며 NTD의 histidine 잔기 2개를 이용하여 공동 상단에서 코발트 이온과 상호작용한다. 또한 구조 비교 분석 결과 bssSirB는 공동 내에 SHC 분자를 수용하는 것으로 판단된다. 이러한 구조적 발견에 기초하여 bssSirB의 공동은 SHC의 코발트 삽입이 이뤄지는 활성 부위임을 제안한다.

• BMB Reports
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• 제43권7호
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• pp.485-490
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• 2010

Neurotrophins regulate many aspects of neuronal function through activation of the high affinity Trk receptors. Shc family proteins are implicated in the coupling of RTK to the Ras/mitogen-activated protein kinase signaling cascade. Here we report that the fourth Shc family member, ShcD, associates with TrkB receptor and regulates BDNF-induced MAPK activation. Yeast two-hybrid assay and Co-IP experiments demonstrate ShcD interacts with TrkB in a kinase-activity-dependent manner. Confocal analysis shows ShcD cololizes well with TrkB in transfected 293T cells. Subsequent mapping experiments and mutational analysis indicate that both PTB and SH2 domains are capable of binding to TrkB and PTB domain binds to TrkB NPQY motif. Furthermore, ShcD is involved in BDNF-induced MAPK activation. In summary, we demonstrate that ShcD is a substrate of TrkB and mediates TrkB downstream signaling pathway.

• Steel and Composite Structures
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• 제34권2호
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• pp.299-308
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• 2020

This study aims to carry out the experimental and numerical investigation on the flexural behavior of sandwich honeycomb composite (SHC) beams reinforced with novel triaxially woven fabric composite skins. Different stacking sequences of the carbon fiber reinforcement polymer (CFRP) laminate; i.e., 0°-direction of TW (TW0), 0°-direction of UD (UD0), and 90°-direction of UD (UD90) were studied, from which the flexural behavior of SHC beam behaviors reinforced with TW0/UD0 or TW0/UD90 novel laminated skins were compared with those reinforced with UD0/90 conventional laminated skins under four-point loading. Generally, TW0/UD0 SHC beams displayed the same flexural stiffness as UD0/90 SHC beams in terms of load-deflection relationships. In contrast, TW0/UD90 SHC beams showed a 70% lower efficiency than those of UD0/90 SHC. Hence, the TW0/UD0 laminate arrangement is more effective with a mass reduction of 39% compared with UD0/90 for SHC beams, although their stiffness and shear strength are practically identical.

• Toxicological Research
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• 제15권1호
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• pp.89-93
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• 1999

2,3,7,8-Tetrachlorodibenzo-p-dioxin(TCDD) is known to induce cytochrome p450 1A1 and to activate c-Src kinase and p21 Ras. This study examined the molecular interactions of adaptor proteins including Shc, Grb2, and Sos in rat primary hepatocytes and their relationship to the induction of CYP1A1 by TCDD. TCDD induced CYP1A1 level and EROD activity in a dose-dependent mode. Sos/Grb2 association isincreased by TCDDㅑㅜ a dose dependent mode. Tyrosine phosphorylated Shc, mainly p152, onloads to Grb2/Sos complex upon TCDD stimulation. The electrophoretic mobility shift of Sos is showed by TCDD. These results indicate that TCDD modulated the molecular interaction features of adaptor compoes proteins including Shc, Grb2, and Cnl in early signaling pathway of TCDD-mediated CYP 1A1 induction of rat primary hepatocyte.

• BMB Reports
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• 제32권2호
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• pp.119-126
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• 1999

The SH3 domain of PLC-${\gamma}1$ has been known to induce DNA synthesis. However, little is known about the putative effector proteins that associate with the domain. In this report, we provide evidence that the SH3 domain of PLC-${\gamma}1$ associates with Shc, which has been implicated in the activation of p21Ras in response to many growth factors. The association between Shc and PLC-${\gamma}1$ is enhanced either by v-Src-induced transformation or EGF-stimulation in vivo and in vitro. Furthermore, from transient expression studies with COS-7 cells, we show that the SH3 domain of PLC-${\gamma}1$ is required for association with Shc in vivo, whereas tyrosyl phosphorylation of PLC-${\gamma}1$ is not. Taken together, we suggest that Shc might be involved in the PLC-${\gamma}1$-mediated signaling pathway.

• 한국의류산업학회지
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• 제22권5호
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• pp.584-594
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• 2020

Growth in the second-hand clothing (SHC) market has become noteworthy, especially for millennial consumers in Korea. This study identifies the moderating role of fashion leadership influencing the relationship between the perceived environmental benefits of SHC and continuance usage intention for millennial consumers. The hypotheses were statistically tested using online survey data, and the respondents were Korean female millennials aged between 25 and 39 years who had online shopping experiences of SHC within 3 months. Furthermore, 263 responses were analyzed by confirmatory factor, hierarchical regression, and conditional process analyses using SPSS, AMOS, and PROCESS v3.3. Results validated that the perceived environmental benefits of SHC and fashion leadership have positive effects on continuance usage intention of purchased second-hand items. Fashion leadership also moderates the relationship between the environmental benefits of SHC and continuance usage intention, showing that as the level of fashion leadership increases, the environmental benefits have more positive effects on continuance usage intention. Theoretical and practical implications were also discussed. This study will help bridge theoretical and practical gaps between purchasing and using SHC by focusing on the interaction effect of fashion leadership and its perceived environmental benefits.