• 제목/요약/키워드: O-GlcNAc

검색결과 38건 처리시간 0.03초

Bacillus subtilis JK-56이 생산하는 chitinase isozyme의 정제와 특성 규명 (Purification and characterization of the chitinase from Bacillus subtilis JK-56)

  • 전홍기;김낙원;정영기
    • 생명과학회지
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    • 제12권1호
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    • pp.77-86
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    • 2002
  • 토양으로부터 chitinase를 생성하는 균주를 분리하여 동정한 결과 Bacillus subtilis로 판명되었으며, 분리한 균주를 Bacillus subtilis JK-56이라 명명하였다. B. subtilis JK-56의 chitinase 생산 최적 조건을 검토한 결과 1% chitin, 0.5% polypeptone, 0.1% KCI, 0.05% MnS $O_4$.4$H_2O$이며 초발 pH 7.0, 배양온도 37$^{\circ}C$에서 가장 많은 효소를 생산하였다. 본 균주가 생산하는 chitinase를 정제하기 위해서 native-PAGE를 이용해 효소활성 band를 확인한 결과, 1개의 강한 활성 band와 2개의 약한 활성 band를 가지는 isozyme으로 확인되었다. 확인된 isozyme을 정제한 결과, isozyme 중 1개의 강한 활성 band를 정제하였고 정제된 효소를 Chi-56A라고 명명하였다 Chi-56A의 효소 특성에 관해서 실험한 결과 분자량은 약 53kDa, pI는 4.3으로 확인되었다. 본 효소는 $65^{\circ}C$까지 상당히 안정하였으며 효소의 최대활성 온도도 $65^{\circ}C$로 확인되는 등 열에 대해 상당히 안정한 효소로 확인되었다. Collidal chitin에 대한 정제효소 Chi-56A의 $K_{m}$ 값은 17.33g/L였다. 그리고 pH 6.0에서 최대의 활성을 나타내었고, 산성범위보다 알칼리범위에서 안정한 것으로 나타났다. 또한 $Mn^{2+}$ 존재 하에서 높은 활성을 나타내었고 C $O^{2+}$$Mg^{2+}$ 존재 하에서도 활성이 약간 증가한 반면에 H $g^{2+}$ 존재 하에서는 상당한 저해를 받았다. Chito 올리고당에 대한 분해 산물을 HPLC로 확인해 본 결과 짝수개의 올리고당의 분해산물은 (GlcNAc)$_2$만을 생산하였고 홀수개의 올리고당에 대해서는 GlcNAc와 (GlcNAc)$_2$를 생산하는 것으로 비환원성 말단으로부터 이당체인 diacetyl chitobiose ((GlcNAc)$_2$)를 생산하는 exo형 chitinase로 추정 된다.

Developmental Patterns of Gal$\beta$1,3(4)GlcNAc $\alpha$2,3-Sialyltransferase (ST3Gal III) Expression in the Mouse: In Situ Hybridization Using DIG-labeled RNA Probes

  • Ji, Min-Young;Lee, Young-Choon;Kim, Kyoung-Sook;Cho, Jin-Won;Jung, Kyu-Yong;Kim, Cheorl-Ho;Choo, Young-Kug
    • Archives of Pharmacal Research
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    • 제22권3호
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    • pp.243-248
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    • 1999
  • Sialic acids are key determinants for biological processes, such as cell-cell interaction and differentiation. Sialyltransferases contribute to the diversity in carbohydrate structure through their attachment of sialic acid in various terminal positions on glycolipid and glycoprotein (N-linked and O-linked) carbohydrate groups. Gal$\beta$ 1,3(4)GlcNAc $\alpha$2,3-sialyltransferase (ST3Gal III) is involved in the biosynthesis of $sLe^{X}$ and sLe^{a}$ known as selection ligands and tumor-associated carbohydrate structures. The appearance and differential distribution of ST3Gal III mRNA during mice embryogenesis [embryonic (E) days; E9, E11, E13, E15] were investigated by in situ hybridization with digoxigenin-labeled RNA probes coupled with alkaline phosphatase detection. On E9, all tissues were positive for ST3Gal III mRNA expression whereas ST3Gal III mRNA on E11 was not detected throughout all tissues. On E13, ST3GAl III mRNA was expressed in different manner in various tissues. In this stage, ST3Gal III mRNA was positive only in the liver, pancreas and bladder. On E15, specific signal for ST3GAl III was detected in the liver, lung and forebrain. These results indicate that ST3Gal III is differently expressed at developmental stages of mice embryo, and this may be importantly related with regulation of organogenesis in mice.

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O-GlcNAcylation of NF-κB Promotes Lung Metastasis of Cervical Cancer Cells via Upregulation of CXCR4 Expression

  • Ali, Akhtar;Kim, Sung Hwan;Kim, Min Jun;Choi, Mee Young;Kang, Sang Soo;Cho, Gyeong Jae;Kim, Yoon Sook;Choi, Jun-Young;Choi, Wan Sung
    • Molecules and Cells
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    • 제40권7호
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    • pp.476-484
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    • 2017
  • C-X-C chemokine receptor 4 (CXCR4) stimulates cancer metastasis. NF-${\kappa}B$ regulates CXCR4 expression in cancer cells, and O-GlcNAc modification of NF-${\kappa}B$ promotes its transcriptional activity. Here, we determined whether CXCR4 expression is affected by O-GlcNAcylation of NF-${\kappa}B$ in lung metastasis of cervical cancer. We found elevated levels of O-linked-N-actylglucosamine transferase (OGT) and O-GlcNAcylation in cervical cancer cells compared to those in non-malignant epithelial cells and detected increased expression of NF-${\kappa}B$ p65 (p65) and CXCR4 in cervical cancer cells. Knockdown of OGT inhibited the O-GlcNAcylation of p65 and decreased CXCR4 expression levels in HeLa cells. Thiamet G treatment increased O-GlcNAcylated p65, which subsequently enhanced CXCR4 expression levels. Inhibition of O-GlcNAcylation by 6-Diazo-5-oxo-L-norleucine (DON) treatment decreased p65 activation, eventually inhibiting CXCR4 expression in HeLa cells. Lung tissues from mice engrafted with OGT-knockdown HeLa cells (shOGT) exhibited lower expression of Ki-67 and HPV E6 and E7 oncogenes compared to lung tissues from mice engrafted with control HeLa cells (shCTL). In addition, lung tissues from mice engrafted with shOGT cells exhibited lower p65 and CXCR4 immunoreactivity compared to tissues from mice engrafted with shCTL cells. Taken together, our data suggest that p65 O-GlcNAcylation promotes lung metastasis of cervical cancer cells by activating CXCR4 expression.

과일 및 야채의 Carbamate계 농약의 잔류량 (Studies on the Residues of Carbamate Pesticide in Fruits and Vegetables)

  • 이해금;최용순;최규열
    • 한국식품위생안전성학회지
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    • 제2권1호
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    • pp.3-8
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    • 1987
  • 춘천시 근교에서 생산된 '84년산 딸기, 토마토, 오이, 포도, 사과, 그리고 배추 각 5점씩 30점에 대한 BPMC, MIPC, NAC의 잔류수준을 분석한 결과 아래와 같은 결론을 얻었다. 1. BPMC의 잔류함량은 딸기에 있어서 평균 0.0243ppm, 토마토 0.0394ppm, 오이 0.0459ppm, 포도 0.0472ppm, 사과 0.0680ppm, 배추 0.0986ppm 수준으로 대상농약중 가장 높은 잔류함량을 보였다. 2. NAC는 포도에서 평균 0.0071ppm, 오이 0.0009ppm, 배추 0.0009ppm 수준으로 검출되었다. 3. MIPC는 배추에서만 평균 0.0017pppm의 낮은 수준으로 보였다. 4. 잔류농약 검출율은 BPMC(80%)>NAC(16.7%)>MIPC(10%) 순이었다. 5. 계절적으로 수확기가 늦은 농산물일수록 비교적 높은 잔류수준을 보였다.

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Glucosamine increases vascular contraction through activation of RhoA/Rho kinase pathway in isolated rat aorta

  • Kim, Do-Hyung;Seok, Young-Mi;Kim, In-Kyeom;Lee, In-Kyu;Jeong, Seong-Yun;Jeoung, Nam-Ho
    • BMB Reports
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    • 제44권6호
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    • pp.415-420
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    • 2011
  • Diabetes is a well-known independent risk factor for vascular disease. However, its underlying mechanism remains unclear. It has been reported that increased influx of the hexosamine biosynthesis pathway (HBP) induces O-GlcNAcylation of proteins, leading to insulin resistance. In this study, we determined whether or not O-GlcNAc modification of proteins could increase vessel contraction. Using an endothelium-denuded aortic ring, we observed that glucosamine induced OGlcNAcylation of proteins and augmented vessel contraction stimulated by U46619, a thromboxane $A_2$ agonist, via augmentation of the phosphorylation of MLC20$MLC_{20}$, MYPT1(Thr855), and CPI17, but not phenylephrine. Pretreatment with OGT inhibitor significantly ameliorated glucosamine-induced vessel constriction. Glucosamine treatment also increased RhoA activity, which was also attenuated by OGT inhibitor. In conclusion, glucosamine, a product of glucose influx via the HBP in a diabetic state, increases vascular contraction, at least in part, through activation of the RhoA/Rho kinase pathway, which may be due to O-GlcNAcylation.