• 제목/요약/키워드: protein bound-SH

검색결과 18건 처리시간 0.027초

노화촉진 생쥐에서 노화에 따른 활성산소 생성 및 항산화능의 변화 (Age-associated Alteration in the Hapatic Superoxide Generation and Antioxidant Activities in the Senescence-accelerated Mice)

  • 정해영;김윤경
    • 약학회지
    • /
    • 제36권5호
    • /
    • pp.460-468
    • /
    • 1992
  • Several bichemical parameters related to free radicals were estimated in senile-prone (P) and resistant(R) strains of male senescence-accelerated mice(SAM) at 2, 5 and 11 months of age. The superoxide generation was increased with age in SAM-R/1 and SAM-P/2. Compared to SAM-R/1, more generation of superoxide was significantly noted in the SAM-P/2 liver. The activities of Cu/Zn-superoxide dismutase and catalase were decreased during aging and these activities in SAM-P/2 were significantly lower than in SAM-R/l liver. The activities of glutathione S-transferase were varied with aging, whereas SAM-P/2 showed lower levels compared to SAM-R/l. The gradual decreases of glutathione, protein bound-SH and nonprotein bound-SH contents were noted with increasing age. SAM-P/2 liver contained lesser amounts of glutathione and nonprotein bound-SH compared to SAM-R. In conclusion, superoxide generation was increased whereas the antioxidant enzyme activities were decreased during aging in SAM-R/1. In addition, SAM-P/2 strain showed more superoxide generation and less antioxidant enzyme activities than SAM-R/1 in the liver, thus we assume that these factors might accelerate the senescence of SAM-P/2 strain.

  • PDF

괄루근으로부터 추출한 다당류의 항당뇨활성 및 당뇨성 쥐의 글루타치온대사에 미치는 영향 (Effect of Polysaccharide from Trichosanthes kirilowii on Antidiabetic Activity and Glutathione Metabolism in Hyperglycemic Rats)

  • 정연봉;이종철
    • 약학회지
    • /
    • 제39권5호
    • /
    • pp.528-534
    • /
    • 1995
  • This investigation was aimed at the study of the antidiabetic activity and effect on hepatic glutathione metabolism of polysaccharide from Trichosanthes kirilowii in hyperglycemic rats with aboxan (175 mg/Kg, i.p.). As the results, the polysaccharide inhibited the increase of blood glucose, triglyceride level and lactate dehydrogenase activity, but cholesterol not changed. And it increased protein bound-SH, nonprotein bound-SH, glutathione level and inhibited the decrease of glutathione S-transferase.

  • PDF

Development of ELISA System for Screening of Specific Binding Inhibitors for Src Homology (SH)2 Domain and Phosphotyrosine Interactions

  • Lee, Sang-Seop;Lee, Kyung-Im;Yoo, Ji-Yun;Jeong, Moon-Jin;Park, Young-Mee;Kwon, Byoung-Mog;Bae, Yun-Soo;Han, Mi-Young
    • BMB Reports
    • /
    • 제34권6호
    • /
    • pp.537-543
    • /
    • 2001
  • In the present study, an in vitro ELISA system to assess the interaction between Src homology (SH)2 domains and phosphotyrosine that contain peptides was established using purified GST-conjugated SH2 proteins and synthetic biotinylated phosphotyrosine that contain oligopeptides. The SH2 domains bound the relevant phosphopeptides that were immobilized in the streptavidin-coated microtiter plate in a highly specific and dose-dependent manner. The epidermal growth factor receptor (EGFR)-, T antigen (T Ag)-, and platelet-derived growth factor receptor (PDGFR)-derived phosphopeptides interacted with the growth factor receptor binding protein (Grb)2/SH2, Lck/SH2, and phosphatidyl inositol 3-kinase (PI3K) p85/SH2, respectively. No cross-reactions were observed. Competitive inhibition experiments showed that a short phosphopeptide of only four amino acids was long enough to determine the binding specificity. Optimal concentrations of the GST-SH2 fusion protein and phosphopeptide in this new ELISA system for screening the binding blockers were chosen at 2nM and 500nM, respectively. When two candidate compounds were tested in our ELISA system, they specifically inhibited the Lck/SH2 and/or p85/SH2 binding to the relevant phosphopeptides. Our results indicate that this ELISA system could be used as an easy screening method for the discovery of specific binding blockers of protein-protein interactions via SH2 domains.

  • PDF

Adriamycin의 독성 발현에 미치는 백렴(Ampelopsis radix)의 영향 (Effect of Ampelopsis radix on the Toxicity of Adriamycin)

  • 김동석;이성호;정연봉
    • 한국식품영양학회지
    • /
    • 제7권3호
    • /
    • pp.232-238
    • /
    • 1994
  • Adriamycin is a major cancer chemotherapeutic agent against a me range of human neoplasms. However, its clinical application is limited since It has a variety of side effects, bone marrow suppression and cardiotoxity, and this toxicity appears by free radical. This study investigated the effect of Ampelopsis radix on the toxicity of adriamycin. The methanol fraction reduced slightly adriamycin induced lipid peroxidation and superoxide production at the dose of 50mg /kg. 1.p., respectively. During the adriamycin administration. Protein bound-SH, nonprotein bound-SH, and glutathione-5-transferase did not change, but methanol fraction treated group were markedly increase. These results indicated that Ampelopsis radix has a major influence on the thiol group and related enzyme activity on the antioxidative effects.

  • PDF

저근백피(樗根白皮) 성분(成分)의 생리활성(生理活性)에 관한 연구(硏究) (II) -클로로포름분획의 급성 및 신장에 대한 독성- (Studies on the Biologic Activities of the Constituents of Ailanthi Cortex Radicis (II) -Acute and Renal Toxicity of Chloroform Fraction-)

  • 김종;김혜경;박수완;최종원;이정규
    • 생약학회지
    • /
    • 제25권2호
    • /
    • pp.140-143
    • /
    • 1994
  • During the serial attempts to identify the chemical and biological characteristics of Ailanthi Cortex Radicis, the root bark of Ailanthus altissima (Simaroubaceae), we find out the serious toxic effect on kidney by chloroform fraction of the methanolic extract of the herb drug. The toxicities were revealed as the increase of urea nitrogen amount in blood and lactate dehydrogenase and ${\gamma}-glutamyltransferase$ activities in urine and the decrease of the concentration of glutathione and both of protein bound and non-protein bound -SH in kidney tissue.

  • PDF

Solution Dynamics Studies for the Lck SH2 Domain Complexed with Peptide and Peptide-Free Forms

  • Yoon, Jeong-Hyeok;Chi, Myung-Whan;Yoon, Chang-No;Park, Jongsei
    • 한국응용약물학회:학술대회논문집
    • /
    • 한국응용약물학회 1995년도 춘계학술대회
    • /
    • pp.81-81
    • /
    • 1995
  • It is well known that Src Homology 2(SH2) domain in many intracellular signal transduction proteins is very important. The domain has about 100 amino acid residues and bind phosphotyrosine-containing peptide with high affinity and specificity. Lck SH2 domain is a Src-like, lymphocyte-specific tyrosine kinase. An 11-residue phosphopeptide derived from the hamster polvoma middle-T antigen, EPQp YEEIPIYL, binds with an 1 nM dissociation constant to Lck SH2 domain. And it is known that the phosphotyrosine and isoleucine residues of the peptide are tightly bound by two well-defined pockets on Lck SH2 domain's surface. To investigate the conformational changes during complexation of SH2 domain with phosphopeptide we have performed the molecular dynamics simulation for Lck SH2 domain with peptide and peptide-free form at look in aqueous solution. More than 3000 water molecules were incorporated to solvate Lck SH2 domain and peptide. Periodic boundary condition has been applied in molecular dynamics simulation. Data analysis with the results of that simulation shows that the phosphopeptide makes primary interaction with the Lck SH2 domain at six central residues, The comparison of the complexed and uncomplexed SH2 domain structures in solution has revealed only relatively small change. But the hydrophilic and hydrophobic pockets in the protein surface show the conformational changes in spite of the small structural difference between the complex and peptide-free forms.

  • PDF

노화과정(老化過程)의 흰쥐에서 보신환(補腎丸)이 신장(腎臟)의 대사효소계(代謝酵素系)에 미치는 영향(影響) (Effect of BOSINHWAN(BSH) Water Extract on Renal Lipid Peroxide Content and Metabolic Enzyme System)

  • 손민성;오민석;송태원
    • 혜화의학회지
    • /
    • 제8권1호
    • /
    • pp.659-674
    • /
    • 1999
  • The experimental studies were carried out in order to prove the effect of BSH water extract on Renal lipid peroxide content and metabolic enzyme system experimental studies about peroxide content, transferase, enzyme activity were carried out. The result were obtained as follows : 1. In the change of lipid peroxide of renal tissue, all group was decreased, more of two weeks was decreased. 2. In the Change of BUN of renal tissue, all group was decreased. 3. In the change of LDH of urine, all group was not significant. 4. In the change of ${\gamma}$-glutamyltransferasde, Xanthine oxidase, Aldehyde oxidase of urine, all group was decreased. 5. In the change of protein-bound SH, nonprotein-bound SH, glutathione, glutathione S-transferase, ${\gamma}$-Glutamylcystein synthetase of renal tissue, all group was increased. From above results, BSH was had significant effects on the senile, so it is expected to clinical application on senility and geratology.

  • PDF

The Mitochondrial Tricarboxylate Carrier of Silver Eel: Chemical Modification by Sulfhydryl Reagents

  • Capobianco, Loredana;Impagnatiello, Tecla;Ferramosca, Alessandra;Zara, Vincenzo
    • BMB Reports
    • /
    • 제37권5호
    • /
    • pp.515-521
    • /
    • 2004
  • The tricarboxylate (or citrate) carrier was purified from eel liver mitochondria and functionally reconstituted into liposomes. Incubation of the proteoliposomes with various sulfhydryl reagents led to inhibition of the reconstituted citrate transport activity. Preincubation of the proteoliposomes with reversible SH reagents, such as mercurials and methanethiosulfonates, protected the eel liver tricarboxylate carrier against inactivation by the irreversible reagent N-(1-pyrenyl)maleimide (PM). Citrate and L-malate, two substrates of the tricarboxylate carrier, protected the protein against inactivation by sulfhydryl reagents and decreased the fluorescent PM bound to the purified protein. These results suggest that the eel liver tricarboxylate carrier requires a single population of free cysteine(s) in order to manifest catalytic activity. The reactive cysteine(s) is most probably located at or near the substrate binding site of the carrier protein.

Association of Killer Cell Ig-like Receptor (KIR) with an Adaptor Protein Shc

  • Cho, Hyun-Il;Chwae, Yong-Joon;Park, Sang-Myun;Kim, Jong-Sun
    • IMMUNE NETWORK
    • /
    • 제6권2호
    • /
    • pp.67-75
    • /
    • 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.

Ras-GTPase-activating protein SH3 domain-binding proteins 2, G3BP2와 KIF5A C-말단 꼬리 영역과의 결합 (Interaction of Ras-GTPase-activating Protein SH3 Domain-binding Proteins 2, G3BP2, With the C-terminal Tail Region of KIF5A)

  • 정영주;장원희;이원희;김무성;김상진;엄상화;문일수;석대현
    • 생명과학회지
    • /
    • 제27권10호
    • /
    • pp.1191-1198
    • /
    • 2017
  • 세포 내 소기관들과 소포들은 세포 내에서 미세소관을 따라 적절한 구획으로 수송된다. 이러한 세포 내 수송과정은 분자 모터단백질인 kinesin과 dynein에 의하여 이루어진다. Kinesin 1은 오징어 축삭돌기 세포질로부터 처음 분리되었으며 2개의 중쇄단위체(KHCs, 또는 KIF5s) 및 이와 결합하는 경쇄단위체(KLCs)의 복합체를 형성한다. KIF5s는C-말단 고리 영역을 통해 많은 다양한 단백질과 결합하는데, 아직 그 결합단백질들은 충분히 밝혀지지 않았다. 본 연구에서는 KIF5A 결합단백질을 분리하기 위하여 효모 two-hybrid 탐색을 수행하여 스트레스 과립형성과 mRNP 위치결정에 관여하는 Ras-GTPase-activating protein (GAP) Src homology3 (SH3)-domain-binding protein 2 (G3BP2)를 분리하였다. G3BP2는 KIF5A의 C-말단 고리 영역에 존재하는 73개 아미노산을 포함하는 영역과 결합하였다. 그러나 G3BP2는 KIF5B, KIF5C, KLC1, KIF3A와는 결합하지 않았다. KIF5A는 G3BP2의 arginine-glycine-glycine(RGG)/Gly-rich 도메인과 결합하지만 G3BP1과는 결합하지 않았다. HEK-293T세포에 G3BP2와 KIF5A를 발현하여 면역침강한 결과 G3BP2와 KIF5A는 같이 침강하였다. 또한 HEK-293T 세포 내의 전체에서 두 단백질은 같은 부위에 존재하였다. 이러한 결과들은 세포 내에서 G3BP2는 KIF5A와 결합하는 결합단백질로 확인 되었다.