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The role of residues Cys10 and His106 in glutathione transferase from Escherichia coli

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The role of residues Cys10 and His106 in glutathione transfe...

Characterisation of the activity and folding of the glutathione transferase from Escherichia coli and the role of residues Cys 10 and His 106

第二届全国“跨学科蛋白质研究”学术讨论会

作者：王新宇柯莎 National Laboratory of Biomacromolecules Institute of BiophysicsChinese Academy of Sciences Department of Biophysics Institute of PhysicsNankai University

glutathione transferases (GSTs) are an important class of enzymes involved in cellular *** are found in all classes of organisms and are implicated in resistance towards drugs,pesticides,herbicides and *** activity, structure and folding,particularly of eukaryotic GSTs,has therefore been widely *** crystal structure of GST from Escherichia coli (EGST) was reported around 10 years ago,suggesting Cys 10 and His 106 as potential catalytic residues. However,the role of these residues in catalysis has not been further investigated,nor have the folding properties of the protein been *** this study,we investigate the contributions of residues Cys 10 and His 106 to the activity and stability of *** found that EGST shows a complex equilibrium unfolding profile,involving population of at least 2 partially-folded intermediates,one of which is *** of residues Cys 10 and His 106 leads to stabilization of the protein and affects the apparent steady-state kinetic parameters for enzyme *** results suggest that the aromatic ring of His 106 plays an important role in the catalytic mechanism of the enzyme,while Cys 10 is involved in binding of the substrate *** study demonstrates that protein engineering of the Cys 10 site can increase both the stability and activity of EGST.

关键词： glutathione transferase GST protein structure protein folding enzyme activity

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Characterisation of the activity and folding of the glutathi...

Kinases and glutathione transferases:selective and sensitive targeting

第九届全国酶学学术讨论会暨邹承鲁诞辰85周年纪念会

作者： Sarah Perrett National Laboratory of Biomacromolecules Institute of BiophysicsChinese Academy of Sciences

关键词： glutathione transferase GST protein structure protein folding enzyme activity

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Frontiers in Biology 2011年第2期6卷 156-169页

作者： Yasemin G.ISGOR Belgin S.ISGOR Chemistry Group Faculty of EngineeringAtilim UniversityAnkara 06836Turkey

Kinases,representing almost 500 proteins in the human genome,are responsible for catalyzing the phosphorylation reaction of amino acid residues at their *** the largest family of kinases,the protein tyrosine kinases(PTKs)have roles in controlling the essential cellular activities,and their deregulation is generally related to pathologic *** recent efforts on identifying their signal transducer or mediator role in cellular signaling revealed the interaction of PTKs with numerous enzymes of different classes,such as Ser/Thr kinases(STKs),glutathione transferases(GSTs),and receptor tyrosine kinases(RTKs).In either regulation or enhancing the signaling,PTKs are determined in close interaction with these enzymes,under specific cellular conditions,such as oxidative stress and *** this concept,intensive research on thiol metabolizing enzymes recently showed their involvement in the physiologic functions in cellular signaling besides their well known traditional role in antioxidant *** shared signaling components between PTK and GST family enzymes will be discussed in depth in this research review to evaluate the results of recent studies important in drug targeting for therapeutic intervention,such as cell viability,migration,differentiation and proliferation.

关键词： glutathione transferase protein tyrosine kinase small molecule inhibitors c-Src signal transduction drug targeting

Purification and characterization of rat testicular glutathione S-transferases:role in the synthesis of eicosanoids

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Asian Journal of Andrology 2000年第4期2卷 277-282页

作者： D.Anuradha K.VeeraReddy T.CharlesKumar S.Neeraja P.R.K.Reddy P.Reddanna School of Life Sciences University of Hyderabad Hyderabad-500 046 IndiaSchool of Life Sciences University of Hyderabad Hyderabad-500 046 IndiaSchool of Life Sciences University of Hyderabad Hyderabad-500 046 IndiaSchool of Life Sciences University of Hyderabad Hyderabad-500 046 IndiaSchool of Life Sciences University of Hyderabad Hyderabad-500 046 IndiaSchool of Life Sciences University of Hyderabad Hyderabad-500 046 India

Aim: Purification of glutathione S-transferases (GSTs) from rat testis; separation and identificationunits and their role in eicosanoid biosynthesis. Methods: Purification of mt testicular GSTs by affit?phy, employing S-hexylglutathione-linked epoxy-activated Sepharose 6B column and separation of indiby reverse phase-high pressure liquid chromatography (RP-HPLC). Characterization of affinity purified,um dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Westem blot analysis. The roGSTs in eicosanoid biosynthesis was determined by incubating GSTs with 5, 6-Leukotriene A_4Me (prostaglandin H_2 (PGH_2) and analyzing the products formed on HPLC/TLC. Results: The present stumajority of rat testicular GSTs are of Y_b size (60%) with molecular weight of 27 kDa. The most preunits, however, are GST Y_(n2) (27% ), followed by GST Y_c (24% ) and GST Y_(nl) (20%). These testiculavery high Leukotriene C_4 (LTC_4) synthase activity with 5, 6-Leukotriene A4Me (LTA_4Me) as theprostaglandin D (PGD) synthase activity with prostaglandin H_2 (PGH_2) as the substrate. Conclusion:testicular GSTs are Y_b sized and are involved in the synthesis of eicosanoids like LTC_4 and PGD_2.(Asian J Androl 2000 Dec; 2: 277-282)

关键词： testis glutathione transferase leukotrienes prostaglandins

glutathione S-transferases M1,T1 genotypes and the risk of gastric cancer:A case-control study

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World Journal of Gastroenterology 2001年第4期7卷 506-509页

作者： Lin Cai Shun-Zhang Yu Zuo-Feng Zhang Department of Epidemiology.Fujian Medical University,Fuzhou 350004,Fujian Province,ChinaDepartment of Epidemiology,Shanghai Medical University,Shanghai 200032,China Department of Epidemiology,UCLA School of Public Health,Los Angeles California,USA Department of Epidemiology Fujian Medical University Fuzhou 350004 Fujian Province China. zjcailin@***

AIM: glutathione S-transferases (GSTs) are involved in the detoxification of many potential carcinogens and appear to play a critical role in the protection from the effects of carcinogens. The contribution of glutathione S-transferases M1 and T1 genotypes to susceptibility to the risk of gastric cancer and their interaction with cigarette smoking are still unclear. The aim of this study was to determine whether there was any relationship between genetic polymorphisms of GSTT1 and GSTT1 and gastric cancer. METHODS: A population based case-control study was carried out in a high-risk area, Changle County, Fujian Province, China. The epidemiological data were collected by a standard questionnaire and blood samples were obtained from 95 incidence gastric cancer cases and 94 healthy controls. A polymerase chain reaction method was used to detect the presence or absence of the GSTT1 and GSTT1 genes in genomic DNA. Logistic regression model was employed in the data analysis. RESULTS: An increase in risk for gastric cancer was found among carriers of GSTT1 null genotype. The adjusted odds ratio (OR) was 2.63 95% Confidence Interval (95% CI) 1.17-5.88, after controlling for age, gender, cigarette smoking, alcohol drinking, and fish sauce intake. The frequency of GSTT1 null genotype in cancer cases (43.16%) was not significantly different from that in controls (50.00%). However, the risk for gastric cancer in those with GSTT1 null and GSTT1 non-null genotype was significantly higher than in those with both GSTT1 and GSTT1 non-null genotype (OR = 2.77, 95% CI 1.15-6.77). Compared with those subjects who never smoked and had normal GSTT1 genotype, ORs were 1.60 (95% CI:0.62-4.19) for never smokers with GSTT1 null type, 2.33 (95% CI 0.88-6.28) for smokers with normal GSTT1, and 8.06 (95% CI 2.83-23.67) for smokers with GSTT1 null type. CONCLUSIONS: GSTT1 gene polymorphisms may be associated with genetic susceptibility of stomach cancer and may modulate tobacco-related carcinoge

关键词： Adult Aged Case-Control Studies Female Genetic Predisposition to Disease Genotype glutathione transferase Humans Male Middle Aged Polymorphism, Genetic Research Support, Non-U.S. Gov't Risk Factors Smoking Stomach Neoplasms

GSTT1,GSTM1 and CYP2E1 genetic polymorphisms in gastric cancer and chronic gastritis in a Brazilian population

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World Journal of Gastroenterology 2004年第9期10卷 1240-1245页

作者： Jucimara Colombo Ana Elizabete Silva Andréa Regina Baptista Rossit Alaor Caetano Aldenis Albaneze Borim Durval Wohnrath Departamento de Biologia UNESP-Campus de So José do Rio Preto-SPBrazil Departamento de Doencas Dermatológicas Infecciosas e ParasitáriasFaculdade de MedicinaFAMERPSo José do Rio PretoSPBrazil Hospital de Base Faculdade de MedicinaFAMERE So José do Rio PretoSPBrazil Fundaco Pio Ⅻ BarretosBrazil

AIM:To test the hypothesis that,in the Southeastern Brazilian population,the GSTT1,GSTM1 and CYP2E1 polymorphisms and putative risk factors are associated with an increased risk for gastric cancer. METHODS:We conducted a study on 100 cases of gastric cancer (GC),100 cases of chronic gastritis (CG),and 150 controls (C).Deletion of the GSTT1 and GSTM1 genes was assessed by multiplex ***2E1/Pst1 genotyping was performed using a PCR-RFLP assay. RESULTS:No relationship between GSTT1/GSTM1 deletion and the c1/c2 genotype of CYP2E1 was observed among the three ***,a significant difference between CG and C was observed,due to a greater number of GSTT1/GSTM1 positive genotypes in the CG *** GSTT1 null genotype occurred more frequently in Negroid subjects,and the GSTM1 null genotype in Caucasians,while the GSTM1 positive genotype was observed mainly in individuals with chronic gastritis infected with H pylori. CONCLUSION:Our findings indicate that there is no obvious relationship between the GSTT1,GSTM1 and CYP2E1 polymorphisms and gastric cancer.

关键词： Polymorphism, Genetic Adolescent Adult Aged Aged, 80 and over Brazil Case-Control Studies Chronic Disease Cytochrome P-450 CYP2E1 Female Gastritis Genotype glutathione transferase Humans Male Middle Aged Research Support, Non-U.S. Gov't Risk Factors Stomach Neoplasms

GSTM1,GSTT1,GSTP1 and CYP1A1 genetic polymorphisms and susceptibility to esophageal cancer in a French population:Different pattern of squamous cell carcinoma and adenocarcinoma

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World Journal of Gastroenterology 2004年第23期10卷 3389-3393页

作者： Ahmed Abbas Karine Delvinquière Mathilde Lechevrel Pierre Lebailly Pascal Gauduchon Guy Launoy Fran ois Sichel GRECAN-EA1772 UFR des Sciences Pharmaceutiques Université de Caen Basse-Normandie et Centre Francois BaclesseAvenue du Général Harris14076 Caen cedex 05France GRECAN-EA1772 Université de Caen Basse-Normandie et Registre des Tumeurs Digestives du Calvados UFR de MédecineAvenue de la C te de Nacre14032 Caen cedexFrance

AIM:To evaluate the association between CYP1A1 and GSTs genetic polymorphisms and susceptibility to esophageal squamous cell carcinoma(SCC)and esophageal adenocarcinoma(ADC)in a high risk area of northwest of France. METHODS:A case-control study was conducted to investigate the genetic polymorphisms of these enzymes (CYPIAI*2C and GSTP1 exon 7 Val alleles,GSTMI*2/*2 and GSTTl *2/*2 null genotypes).A total of 79 esophageal cancer cases and 130 controls were recruited. RESULTS:GSTMI*2/*2 and CYPIAI*IA/*2C genotype frequencies were higher among squamous cell carcinomas at a level dose to statistical significance(OR =1.83,95% CI 0.88-3.83,P=0.11;OR=3.03,95% CI 0.93-9.90,P=0.07, respectively).For GSTP1 polymorphism,no difference was found between controls and cases,whatever their histological *** frequency of GSTT1 deletion was observed in ADC group compared to controls with a statistically significant difference(OR=13.31,95% CI 1.66-106.92,Pglutathione.

关键词： Acyltransferases Adenocarcinoma Adult Aged Aged, 80 and over Carcinoma, Squamous Cell Case-Control Studies Cytochrome P-450 CYP1A1 Esophageal Neoplasms Female France Genetic Predisposition to Disease Genotype glutathione transferase Humans Male Middle Aged Polymorphism, Genetic Research Support, Non-U.S. Gov't Risk Factors

Quantification of mRNA by RT-competitive-PCR and high performance liquid chromatography

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Progress in Natural Science:Materials International 2002年第5期 35-40页

作者： Gary WILLIAMSON Institute of Food Research NR4 7UA Norwich Research Park Nutrition and Consumer Science UK

The use of RT-competitive-PCR with high performance liquid chromatography ( HPLC) detection to quantify the absolute number of mRNA copies in mammalian cells is reported. As an example, the glutathione transferase (GST)-α mRNA in human hep-atoma Hep G2 cells has been estimated. A PCR-generated internal standard was used as a competitor, co-amplified with the GST-α target sequence. The RT-competitive-PCR method was improved by designing target and competitor molecules which differed in only 30 base pairs. This allowed the two sequences to be co-amplified with the same efficiency. This improvement also facilitated a wider ratio to be used than previous methods (target:competitor ratio between 0.2 and 5). Products were baseline separated by HPLC using an ion-exchange column readily quantified at 260 nm. To validate the improved methodology, the effect of a known GST-a inducer, the anticancer drug oltipraz, was shown to induce GST-α mRNA up to 3-fold in Hep G2 cells. The RT-competitive PCR-HPLC method pr

关键词： RT-PCR glutathione transferase oltipraz Hep G2 cell HPLC.

Polymorphism of genes encoding drug-metabolizing and inflammation-related enzymes for susceptibility to cholangiocarcinoma in Thailand

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World Journal of Gastrointestinal Pathophysiology 2023年第2期14卷 21-33页

作者： Gyokukou You Lu Zeng Hideaki Tanaka Emi Ohta Takahiro Fujii Kazuhiko Ohshima Masakazu Tanaka Nobuyuki Hamajima Chutiwan Viwatthanasittiphong Mantana Muangphot Dhiraphol Chenvidhya Adisorn Jedpiyawongse Banchob Sripa Masanao Miwa Satoshi Honjo Department of Bioscience Nagahama Institute of Bio-Science and TechnologyNagahama 526-0829ShigaJapan Division of Neuroimmunology Joint Research Center for Human Retrovirus InfectionKagoshima UniversityKagoshima 890-8544KagoshimaJapan Department of Healthcare Administration Nagoya University Graduate School of MedicineNagoya 466-8550AichiJapan Department of Diagnostic and Interventional Radiology Ubon Cancer CentreUbon Ratchathani 34000Thailand Department of Pathology Ubon Cancer CentreUbon Ratchathani 34000Thailand Department of Surgery Ubon Cancer CentreUbon Ratchathani 34000Thailand Research Division National Cancer InstituteBangkok 10400Thailand Department of Pathology Khon Kaen UniversityKhon Kaen 40002Thailand Department of Paediatirics National Hospital OrganizationFukuoka National HospitalFukuoka 811-1394FukuokaJapan

BACKGROUND Cholangiocarcinoma(CCA)is an intractable cancer,and its incidence in north eastern Thailand is the highest *** with the liver fluke Opisthorchis viverrini(OV)has been associated with CCA ***,animal experiments have suggested that OV alone does not induce CCA,but its combination with a chemical carcinogen like nitrosamine can cause experimentally induced CCA in ***,in humans,other environmental and genetic factors may also be *** To examine relations between risk for CCA and genetic polymorphisms in carcinogenmetabolizing and inflammation-related *** This hospital-based case-control study enrolled 95 case-control pairs matched by age(±5 years)and *** examined relations between risk for CCA and genetic polymorphisms in carcinogenmetabolizing and inflammation-related genes,serum anti-OV,alcohol consumption,and *** of CYP2E1,IL-6(-174 and-634),IL-10(-819),and NF-κB(-94)and their cooccurrence with polymorphisms in the drug-metabolizing enzyme gene GSTT1 or GSTM1 were also *** Although CCA risk was not significantly associated with any single polymorphism,persons with the GSTT1 wild-type and CYP2E1 c1/c2+c2/c2 genotype had an increased risk(OR=3.33,95%CI:1.23-9.00)as compared with persons having the GSTT1 wild-type and CYP2E1 c1/c1 wild *** presence of anti-OV in serum was associated with a 7-to 11-fold increased risk,and smoking level was related to an OR of 1.5-1.8 in multivariable analyses adjusted for each of the seven genetic *** In addition to infection with OV,gene-gene interactions may be considered as one of the risk factors for CCA development.

关键词： Opisthorchis glutathione transferase Cytochrome P-450 CYP2E1 Case-control study