• 제목/요약/키워드: enzymes

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Binding Pattern Elucidation of NNK and NNAL Cigarette Smoke Carcinogens with NER Pathway Enzymes: an Onco-Informatics Study

  • Jamal, Qazi Mohammad Sajid;Dhasmana, Anupam;Lohani, Mohtashim;Firdaus, Sumbul;Ansari, Md Yousuf;Sahoo, Ganesh Chandra;Haque, Shafiul
    • Asian Pacific Journal of Cancer Prevention
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    • 제16권13호
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    • pp.5311-5317
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    • 2015
  • Cigarette smoke derivatives like NNK (4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanone) and NNAL (4-(methylnitrosamino)-1-(3-pyridyl)-1-butan-1-ol) are well-known carcinogens. We analyzed the interaction of enzymes involved in the NER (nucleotide excision repair) pathway with ligands (NNK and NNAL). Binding was characterized for the enzymes sharing equivalent or better interaction as compared to +Ve control. The highest obtained docking energy between NNK and enzymes RAD23A, CCNH, CDK7, and CETN2 were -7.13 kcal/mol, -7.27 kcal/mol, -8.05 kcal/mol and -7.58 kcal/mol respectively. Similarly the highest obtained docking energy between NNAL and enzymes RAD23A, CCNH, CDK7, and CETN2 were -7.46 kcal/mol, -7.94 kcal/mol, -7.83 kcal/mol and -7.67 kcal/mol respectively. In order to find out the effect of NNK and NNAL on enzymes involved in the NER pathway applying protein-protein interaction and protein-complex (i.e. enzymes docked with NNK/NNAL) interaction analysis. It was found that carcinogens are well capable to reduce the normal functioning of genes like RAD23A (HR23A), CCNH, CDK7 and CETN2. In silico analysis indicated loss of functions of these genes and their corresponding enzymes, which possibly might be a cause for alteration of DNA repair pathways leading to damage buildup and finally contributing to cancer formation.

A Stereochemical Aspect of Pyridoxal 5' -Phosphate Dependent Enzyme Reactions and Molecular Evolution

  • Jhee, Kwang-Hwan;Tohru, Yoshimura;Yoichi, Kurokawa;Nobuyoshi, Esaki;Kenji, Soda
    • Journal of Microbiology and Biotechnology
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    • 제9권6호
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    • pp.695-703
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    • 1999
  • We have studied the stereospecificities of various pyridoxal 5'-phosphate (PLP) dependent enzymes for the hydrogen transfer between the C-4' of a bound coenzyme and the C-2 of a substrate in the transamination catalyzed by the enzymes. Stereospecificities reflect the structures of enzyme active-sites, in particular the geometrical relationship between the coenzyme-substrate Schiff base and the active site base participating in an $\alpha$-hydrogen abstraction. The PLP enzymes studied so far catalyze only a si-face specific (pro-S) hydrogen transfer. This stereochemical finding suggests that the PLP enzymes have the same topological active-site structures, and that the PLP enzymes have evolved divergently from a common ancestral protein. However, we found that o-amino acid aminotransferase, branched chain L-amino acid aminotransferase, and 4-amino-4-deoxychorismate lyase, which have significant sequence homology with one another, catalyze a re-face specific (pro-R) hydrogen transfer. We also showed that PLP-dependent amino acid racemases, which have no sequence homology with any aminotransferases, catalyze a non-stereospecific hydrogen transfer: the hydrogen transfer occurs on both faces of the planar intermediate. Crystallographical studies have shown that the catalytic base is situated on the re-face of the C-4' of the bound coenzyme in o-amino acid aminotransferase and branched chain L-amino acid aminotransferase, whereas the catalytic base is situated on the si-face in other aminotransferases (such as L-aspartate aminotransferase) catalyzing the si-face hydrogen transfer. Thus, we have clarified the stereospecificities of PLP enzymes in relation with the primary structures and three-dimensional structures of the enzymes. The characteristic stereospecificities of these enzymes for the hydrogen transfer suggest the convergent evolution of PLP enzymes.

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Structure function relationships amongst the purple acid phosphatase family of binuclear metal-containing enzymes

  • Hamilton, Susan
    • 한국생물정보학회:학술대회논문집
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    • 한국생물정보시스템생물학회 2003년도 제2차 연례학술대회 발표논문집
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    • pp.5-5
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    • 2003
  • The purple acid phosphatases comprise a family of binuclear metal-containing enzymes. The metal centre contains one ferric ion and one divalent metal ion. Spectroscopic studies of the monomeric, ${\sim}$36 kDa mammalian purple acid phosphatases reveal the presence of an Fe(III)Fe(II) centre in which the metals are weakly antiferromagnetically coupled, whereas the dimeric, ${\sim}$110 000 kDa plant enzymes contain either Fe(III)Zn(II) or Fe(III)Mn(II). The three dimensional structures of the red kidney bean and pig enzymes show very similar arrangements of the metal ligands but some significant differences beyond the immediate vicinity of the metals. In addition to the catalytic domain, the plant enzyme contains a second domain of unknown function. A search of sequence databases was undertaken using a sequence pattern which includes the conserved metal-binding residues in the plant and animal enzymes. The search revealed the presence in plants of a 'mammalian-type' low molecular weight purple acid phosphatase, a high molecular weight form in some fungi, and a homologue in some bacteria. The catalytic mechanism of the enzyme has been investigated with a view to understanding the marked difference in specificity between the Fe-Mn sweet potato enzyme, which exhibits highly efficient catalysis towards both activated and unactivated phosphate esters, and other PAPs, which hydrolyse only activated esters. Comparison of the active site structures of the enzymes reveal some interesting differences between them which may account for the difference. The implications fur understanding the physiological functions of the enzymes will be discussed.

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Effect of Superoxide Dismutase and Low Molecular Mediators on Lignin Degradation

  • Leonowicz, Andrzej;Matuszewska, Anna;Luterek, Jolanta;Ziegenhagen, Dirk;Wojtas-Wasilewska, Maria;Hofrichter, Martin;Rogalski, Jerzy;Cho, Nam-Seok
    • Journal of the Korean Wood Science and Technology
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    • 제27권4호
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    • pp.1-14
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    • 1999
  • As the biodegradation of wood constituents has been understood as a multi-basidiomycetes and enzymatic processes, this review will focus on the roles of low molecular compounds and radicals working in harmony with fungal enzymes. Wood rotting basidiomycete fungi penetrate wood, and lead to more easily metabolize carbohydrates of the wood complex. The white-rot fungi, having versatile enzymes, are able to attack directly the "lignin barrier". They also use a multi-enzyme system including so-called "feedback" type enzymes allowing for simultaneous degradation of lignin and carbohydrates. The multi-enzymes including laccase support the proposed route by explaining how the high molecular weight enzymes can function in the wood complex. These enzymes may function separately or cooperate each other. In addition, veratryl alcohol oxidase, cellobiose dehydrogenase, arylalcohol dehydrogenase, and particularly low molecular mediators and radicals have an important role in wood biodegradation. However, the possibility of other mechanism as well as other enzymes, as operating as feedback systems in the process of wood degradation, could not be excluded.

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Hydrolysis of Empty Fruit Bunch of Oil Palm Using Cellulolytic Enzymes from Aspergillus terreus IMI 28243

  • Kader, Jalil;Krishnasamy, Getha;Mohtar, Wan;Omar, Othman
    • Journal of Microbiology and Biotechnology
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    • 제9권4호
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    • pp.514-517
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    • 1999
  • Hydrolysis of EFB (empty fruit bunch) derived from oil palm was studied using crude enzyme from Aspergillus terreus IMI 282743 along with commercial enzymes from Trichoderma reesei and Aspergillus niger. Hydrolysis at $40^{\circ}C$ and $50^{\circ}C$ with $\alpha$-cellulose or EFB gave significantly lower yield when commercial enzymes of T. reesei and A. niger were used and the hydrolysis time extended beyond 10 h. After 24 h of hydrolysis at $40^{\circ}C$ and $50^{\circ}C$, the filter paper activity (Fpase) from A. terreus retained as much activity as A. niger and it was significantly higher than T. reesei. Glucose concentration of 0.25% and 0.5% caused significant inhibition in the crude enzyme, but in regards to the commercial enzymes it only showed a slight effect. Crude enzymes from A. terreus could produce the highest reducing sugars when compared to commercial enzymes from T. reesei or A. niger. Nevertheless, low yield of sugar was observed for EFB for all treatments.

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Inhibitory Effects of Bifidobacterium spp. Isolated from a Healthy Korean on Harmful Enzymes of Human Intestinal Microflora

  • Park, Hye-Young;Bae, Eun-Ah;Han, Myung-Joo;Choi, Eung-Chil;Kim, Dong-Hyun
    • Archives of Pharmacal Research
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    • 제21권1호
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    • pp.54-61
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    • 1998
  • Five hundreds of bifidobacteria were isolated from a healthy Korean and the inhibitory effects of these isloated bacteria on harmful enzymes of human intestinal microflora were examined by cocultivation of the isolated bifidobacteria with E. coli or total human intestinal microflora. In comparison with the results of E. coli or intestinal microflora cultivation, Bifidobacterium breve K-110, B. breve K-111 and B. infantis K-525 effectively inhibited harmful enzymes ($\beta$-glucuronidase and tryptophanase) of E. coli and lowered the pH of the culture media. Also they inhibited the harmful enzymes ($\beta$-glucosidase, $\beta$-glucuronidase, tryptophanase and urease) and ammonia production of intestinal microflora, and lowered pH of the culture media by increasing lactic acid bacteria of intestinal microflora. When these isolated bifidobacteria were administered on mice, fecal harmful enzymes were also inhibited. Among tested bifidobacteria, B. breve K-110 had the highest inhibitory effect of fecal harmful enzymes.

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Purification and Characterization of Two Extracellular Proteases from Oligotropha carboxydovorans DSM 1227

  • Kang, Beom-Sik;Jeon, Sang-Jun;Kim, Young-Min
    • Journal of Microbiology
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    • 제37권1호
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    • pp.14-20
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    • 1999
  • Two extracellular proteases, EP I and EP II, from cells of Oligotropha carboxydovorans (formerly Pseudomonas carboxydovorans) DSM 1227 grown in nutrient broth were purified to greater than 95% homogeneity in five steps using azocasein as a substrate. The final specific activities of EPs I and II were 214.9 and 667.4 units per mg of protein. The molecular weights of native EPs I and II were determined to be 23,000. Sodium dodecyl sulfate-gel electrophoresis revealed the two enzymes to be monomers. The enzymes were found to be serine-type proteases. The activity of EP I was stimulated by Ca2+, Mg2+, and Ba2+, but that of EP II was not. The enzymes were completely inhibited by Fe2+, Hg2+, Co2+, Zn2+, and Cd2+. EDTA and EGTA exhibited a strong inhibitory effect on EP I. The optimal pH for the two enzymes was pH 9.0. The optimal temperatures for EP I and II were 60 and 50$^{\circ}C$, respectively. The enzymes were stable under alkaline conditions. The thermal stability of EP I was higher than that of EP II. Cell-free extracts did not inhibit the purified enzymes. The enzymes were active on casein, azocasein, azocoll, and carbon monoxide dehydrogenase, but weakly active with bovine serum albumin.

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Enzymatic Deinking of Old Newsprit with Alkalophilic Enzymes from Coprinus cinereus 2249

  • Lee, Jung-Myong;Eom, Tae-Jin
    • 펄프종이기술
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    • 제31권5호
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    • pp.12-17
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    • 1999
  • This study was to evaluate applications of the alkalophilic enzymes from Coprinus cinereus 2249 with old newsprint(ONP). A enzymatic deinking process based on alkalopholic enzymes was investigated . It was found that alkalophilic enzymes could effectively deink old newsprint. When applied on deinking of the old newsprint, it increases the freeness and brightness due to effect of hydrolysis at 0.1% enzyme concentration . Also, the physical properties of deinked pulp were improved.

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인삼 Saponin 분획의 고혈당강하작용에 관한 연구(II) (Study on the Hypoglycemic Action of Ginseng Saponin on Streptozotocin Induced Diabetic Rats (II))

  • 주충노;윤수희
    • Journal of Ginseng Research
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    • 제16권3호
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    • pp.198-209
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    • 1992
  • The decreased activities of liver enzymes relating to carbohydrate metabolism such as glucose- 6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase and acetyl CoA carboxylase of streptozotocin injected rats were significantly modified by the intraperitoneal injection of ginseng saponin mixture and/or purified ginsenosides. However, several enzymes such as pyruvate kinase, malic enzyme and glycogen phosphorylase were not modified appreciably by the saponin administration, suggesting that the effect of ginseng saponin might be depend upon individual enzymes. Examination of liver enzymes by liver professing technique using perfusion buffer containing saponin (10-3%) showed that the ginseng saponin might stimulate insulin biosynthesis as well as the related enzyme activities.

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Effects of nandrolone decanoate on expression of steroidogenic enzymes in the rat testis

  • Min, TaeSun;Lee, Ki-Ho
    • Asian-Australasian Journal of Animal Sciences
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    • 제31권5호
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    • pp.658-671
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    • 2018
  • Objective: Nandrolone decanoate (ND) is an anabolic-androgenic steroid frequently used for clinical treatment. However, the inappropriate use of ND results in the reduction of serum testosterone level and sperm production. The suppressive effect of ND on testosterone production has not been investigated in detail. The present study was designed to examine the effect of ND on the expression of steroidogenic enzymes in the rat testis. Methods: Male Sprague Dawley rats at 50 days of age were subcutaneously administrated with either 2 or 10 mg of ND/kg body weight/week for 2 or 12 weeks. The changes of transcript and protein levels of steroidogenic enzymes in the testis were determined by real-time polymerase chain reaction and western blotting analyses, respectively. Moreover, immunohistochemical analysis was employed to determine the changes of immunostaining intensity of these enzymes. The steroidogenic enzymes investigated were steroidogenic acute regulatory protein, cytochrome P450 side chain cleavage enzyme, $17{\alpha}-hydroxylase$, $3{\beta}-hydroxysteroid$ dehydrogenase, and cytochrome P450 aromatase. Results: The treatment of ND resulted in depletion of Leydig cells and sloughing of germ cells in the testis. The ND treatment caused significant expressional decreases of steroidogenic enzymes at transcript and protein levels, and the destructive effects of ND on the testis were more apparent with a higher dose and a longer period of the treatment. Evident reduction of immunostaining intensity present in Leydig cells was clearly detected by the ND treatment. Conclusion: The exposure to ND in young male results not only in histological changes of the testis but also in aberrant gene expression of testicular steroidogenic enzymes, consequently leading into the reduction of testosterone production in the testis and thus likely disruption of spermatogenesis.