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David Mantle - One of the best experts on this subject based on the ideXlab platform.
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In vivo effects of cyfluthrin on proteolytic enzyme activities of malathion-resistant and susceptible strains of Tribolium castaneum
Pakistan Journal of Zoology, 2004Co-Authors: Mushtaq A. Saleem, David Mantle, Richard M. Wilkins, Abdul Rauf ShakooriAbstract:To elucidate whether insecticide toxicity in insects involves insecticide - induced abnormalities of the intracellular protein catabolic process, we have determined the in vivo effect of a synthetic pyrethroid, cyfluthrin on the activities of representative protein catabolising cytoplasmic and lysosomal proteases (responsible for the various stages of the protein degradation cascade and essential for normal cell functioning) in resistant and susceptible strains of Tribolium castaneum. Effect of cyfluthrin was determined at LC 50 after 48 hour of treatment both in the live and dead adult beetles and compared with controls. In treated live beetles, cyfluthrin decreased alanyl Aminopeptidase (18%), arginyl Aminopeptidase (6%), Leucyl Aminopeptidase (22%), tripeptidyl Aminopeptidase (12%), proline endopeptidase (43%) and dipeptidyl Aminopeptidase I (23%) and increased cathepsin H (42%), while in dead beetles, almost all cytoplasmic proteases as well as cathepsin H manifested further decreasing trend. On the other hand in treated resistant strain live beetles, all lysosomal proteases and Leucyl Aminopeptidase were considerably decreased ranging from 19% to 58% of control activity and this decreasing trend was further intensified in treated dead beetles, which ranged from 30% to 96%. We conclude that the effect of cyfluthrin on proteolytic enzyme activities induced inhibition of proteases and could be important in the development of insecticide resistance in T. castaneum.
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Effect of starvation on proteases in insecticide-resistant and susceptible strains of Musca domestica
Pakistan Journal of Zoology, 2003Co-Authors: Mushtaq A. Saleem, David Mantle, Richard M. Wilkins, Sohail Ahmad, Abdul Rauf ShakooriAbstract:To determine the possible involvement of the process of intracellular protein catabolism in the development of insecticide resistance, we have compared the activities of a comprehensive range of cytoplasmic and lysosomal proteolytic enzymes in 48 hours starved and un-starved resistant and susceptible strains of Musca domestica. Compared to un-starved, the homogenates of 48 hours starved house flies of both resistant and susceptible strains showed elevated levels of most of the proteases tested in this study. Thus in susceptible strain, Leucyl Aminopeptidase, proline endopeptidase and cathepsin L showed elevated levels in starved than un-starved by 42%, 28% and 23%, respectively, while the remaining proteases exhibited minor fluctuations. On the other hand, in resistant strain, Leucyl Aminopeptidase (58%), proline endopeptidase (41%) and cathepsin L (38%) showed elevated levels whereas the remaining proteases exhibited minor deviations. The results suggested that higher induction of proteases in starved resistant than susceptible strain may be important in the development of insecticide resistance in M. domestica.
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Effect of starvation on proteases in insecticide-resistant and susceptible strains of Tribolium castaneum
Pakistan Journal of Zoology, 2003Co-Authors: Mushtaq A. Saleem, David Mantle, Richard M. Wilkins, Abdul Rauf ShakooriAbstract:To elucidate the possible involvement of the process of intracellular protein catabolism in the development of insecticide resistance, we have compared the activities of a comprehensive range of cytoplasmic and lysosomal proteolytic enzymes in 48 hours starved and un-starved resistant (CTC-12) and susceptible (FSS-II) strains of red flour beetle, Tribolium castaneum (Herbst.) (Coleoptera: Tenebrionidae) under laboratory conditions. Compared to un-starved, the homogenates of 48 hours starved adult beetles of both resistant and susceptible strains showed elevated levels of all proteases tested in this study. In starved resistant beetles, all proteases except two were considerably increased which included alanyl Aminopeptidase (41%), arginyl Aminopeptidase (24%), Leucyl Aminopeptidase (154%), dipeptidyl Aminopeptidase IV (27%), tripeptidyl Aminopeptidase (26%), proline endopeptidase (316%), dipeptidyl Aminopeptidase I (153%), dipeptidyl Aminopeptidase II (50%), cathepsin B (22%) and cathepsin L (48%). On the other hand, in starved susceptible beetles, only two proteases viz Leucyl Aminopeptidase (34%) and proline endopeptidase (66%) were increased, while the remaing showed less than 20% elevation. The results suggested that more number and higher induction of proteases in starved resistant than susceptible strain beetles may be important in the development of insecticide-resistance in T. caztaneum.
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Comparison of structural protein and proteolytic enzyme levels in degenerating and regenerating rat muscle induced by Notechis scutatus venom.
Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology, 1995Co-Authors: Abul Faiz, John Harris, Charlotte A. Maltin, David MantleAbstract:To develop a clearer understanding of the biochemical mechanism of muscle degeneration and regeneration induced by a single dose of Notechis scutatus scutatus venom, we have correlated changes in the levels of a series of muscle structural proteins and proteolytic enzymes. The degradation of structural proteins post-injection fell into two broad groups; those completely degraded within 3–6 hr (e.g. C- and M-proteins, skelemin), and within 1-2 days (e.g. myosin, actin, troponin), respectively. Similarly, activation of proteases followed two general patterns; those enzymes showing substantially increased activity after 12–24 hr (lysosomal cathepsins, Leucyl Aminopeptidase) and those enzymes showing decreased activity after 12–24 hr, with substantially increased activity after 3–4 days (mainly cytoplasmic proteases). The data suggest that activation of cathepsins B, L and D and in particular Leucyl Aminopeptidase, may be responsible for the early stages of structural protien catabolism, and are thus potential therapeutic targets to prevent myonecrosis following envenomation.
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Biochemical analysis of degeneration and regeneration in rat soleus muscle induced by venom from three subspecies of Daboia russelli
Comparative Biochemistry and Physiology Part B: Comparative Biochemistry, 1994Co-Authors: Abul Faiz, John Harris, David MantleAbstract:To develop a clearer understanding of the biochemical mechanism of muscle degeneration and regeneration induced by a single dose of D. r. pulchella, D. r. russelli or D. r. siamensis venoms, we have correlated changes in the levels of a series of muscle structural proteins and proteolytic enzymes; the sequence of changes were broadly similar, although some differences were noted in relative myotoxicity and regeneration efficiency. The data suggest that activation of cathepsins B, L and D, and Leucyl Aminopeptidase may be of particular importance in the early stages of structural protein degradation, and are thus potential therapeutic targets to prevent myonecrosis following envenomation.
Abdul Rauf Shakoori - One of the best experts on this subject based on the ideXlab platform.
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In vivo effects of lambda-cyhalothrin on proteases of various body compartments of Periplaneta americana adults.
Pakistan Journal of Zoology, 2010Co-Authors: Mushtaq A. Saleem, Richard M. Wilkins, Abdul Rauf ShakooriAbstract:The overall activity and body comportment distribution of a wide range of cytoplasmic and lysosomal proteolytic enzymes was determined in the adults of American cockroach, Periplaneta americana. Individual proteolytic enzymes showed different overall and relative levels of activity in head, thorax, abdomen, leg, internal leg muscle and gut. The gut showed highest activities of alanyl Aminopeptidase, dipeptidyl Aminopeptidase (DAP) I, II and IV, and cathepsin B, L and H while head showed highest activities of arginyl, Leucyl and tripeptidyl Aminopeptidase. In vivo effects of lambda-cyhalothrin on proteases of various body compartments showed that almost all proteases in the gut, except cathepsin B, showed considerably elevated levels ranging from 19% in the case of proline endopeptidase and 429% in the case of DAP I. All DAPs manifested highest levels i.e. DAP I by 429%, DAP II by 355% and DAP IV by 178%, followed by cathepsin L (246%), cathepsin D (123%), tripeptidyl Aminopeptidase (109%), arginyl Aminopeptidase (78%), alanyl Aminopeptidase (72%), Leucyl Aminopeptidase (49%), cathepsin H (37%) and proline endopeptidase (19%). Proteases of all other body compartments exhibited mixed responses, although in head and thorax, all cytoplasmic proteases were increased while almost all lysosomal proteases were decreased.
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In vivo effects of cyfluthrin on proteolytic enzyme activities of malathion-resistant and susceptible strains of Tribolium castaneum
Pakistan Journal of Zoology, 2004Co-Authors: Mushtaq A. Saleem, David Mantle, Richard M. Wilkins, Abdul Rauf ShakooriAbstract:To elucidate whether insecticide toxicity in insects involves insecticide - induced abnormalities of the intracellular protein catabolic process, we have determined the in vivo effect of a synthetic pyrethroid, cyfluthrin on the activities of representative protein catabolising cytoplasmic and lysosomal proteases (responsible for the various stages of the protein degradation cascade and essential for normal cell functioning) in resistant and susceptible strains of Tribolium castaneum. Effect of cyfluthrin was determined at LC 50 after 48 hour of treatment both in the live and dead adult beetles and compared with controls. In treated live beetles, cyfluthrin decreased alanyl Aminopeptidase (18%), arginyl Aminopeptidase (6%), Leucyl Aminopeptidase (22%), tripeptidyl Aminopeptidase (12%), proline endopeptidase (43%) and dipeptidyl Aminopeptidase I (23%) and increased cathepsin H (42%), while in dead beetles, almost all cytoplasmic proteases as well as cathepsin H manifested further decreasing trend. On the other hand in treated resistant strain live beetles, all lysosomal proteases and Leucyl Aminopeptidase were considerably decreased ranging from 19% to 58% of control activity and this decreasing trend was further intensified in treated dead beetles, which ranged from 30% to 96%. We conclude that the effect of cyfluthrin on proteolytic enzyme activities induced inhibition of proteases and could be important in the development of insecticide resistance in T. castaneum.
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Effect of starvation on proteases in insecticide-resistant and susceptible strains of Musca domestica
Pakistan Journal of Zoology, 2003Co-Authors: Mushtaq A. Saleem, David Mantle, Richard M. Wilkins, Sohail Ahmad, Abdul Rauf ShakooriAbstract:To determine the possible involvement of the process of intracellular protein catabolism in the development of insecticide resistance, we have compared the activities of a comprehensive range of cytoplasmic and lysosomal proteolytic enzymes in 48 hours starved and un-starved resistant and susceptible strains of Musca domestica. Compared to un-starved, the homogenates of 48 hours starved house flies of both resistant and susceptible strains showed elevated levels of most of the proteases tested in this study. Thus in susceptible strain, Leucyl Aminopeptidase, proline endopeptidase and cathepsin L showed elevated levels in starved than un-starved by 42%, 28% and 23%, respectively, while the remaining proteases exhibited minor fluctuations. On the other hand, in resistant strain, Leucyl Aminopeptidase (58%), proline endopeptidase (41%) and cathepsin L (38%) showed elevated levels whereas the remaining proteases exhibited minor deviations. The results suggested that higher induction of proteases in starved resistant than susceptible strain may be important in the development of insecticide resistance in M. domestica.
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Effect of starvation on proteases in insecticide-resistant and susceptible strains of Tribolium castaneum
Pakistan Journal of Zoology, 2003Co-Authors: Mushtaq A. Saleem, David Mantle, Richard M. Wilkins, Abdul Rauf ShakooriAbstract:To elucidate the possible involvement of the process of intracellular protein catabolism in the development of insecticide resistance, we have compared the activities of a comprehensive range of cytoplasmic and lysosomal proteolytic enzymes in 48 hours starved and un-starved resistant (CTC-12) and susceptible (FSS-II) strains of red flour beetle, Tribolium castaneum (Herbst.) (Coleoptera: Tenebrionidae) under laboratory conditions. Compared to un-starved, the homogenates of 48 hours starved adult beetles of both resistant and susceptible strains showed elevated levels of all proteases tested in this study. In starved resistant beetles, all proteases except two were considerably increased which included alanyl Aminopeptidase (41%), arginyl Aminopeptidase (24%), Leucyl Aminopeptidase (154%), dipeptidyl Aminopeptidase IV (27%), tripeptidyl Aminopeptidase (26%), proline endopeptidase (316%), dipeptidyl Aminopeptidase I (153%), dipeptidyl Aminopeptidase II (50%), cathepsin B (22%) and cathepsin L (48%). On the other hand, in starved susceptible beetles, only two proteases viz Leucyl Aminopeptidase (34%) and proline endopeptidase (66%) were increased, while the remaing showed less than 20% elevation. The results suggested that more number and higher induction of proteases in starved resistant than susceptible strain beetles may be important in the development of insecticide-resistance in T. caztaneum.
Božić Nataša - One of the best experts on this subject based on the ideXlab platform.
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Characterisation of Leucyl Aminopeptidase from Solanum tuberosum tuber
Elsevier Sci Ltd Oxford, 2010Co-Authors: Vujčić Zoran, Lončar, Nikola L., Dojnov Biljana, Milovanovic Aleksandra, Vujčić Miroslava, Božić NatašaAbstract:Potato juice (a waste product from the starch industry) is a potential source of novel enzymes for food applications. For use in the production and improvement of food protein hydrolysates, commercially available exopeptidases, predominantly Aminopeptidases, are recommended. The present study was performed to explore possible biotechnological interest of Leucyl Aminopeptidase (LAP) activity in the potato tuber. The LAP from potato tuber was purified and characterised. Specific LAP activity was increased 200-fold by purification of the crude extract. The purified enzyme had a pH optimum of 9.0 and temperature optimum of 45 degrees C. LAP hydrolysed leucine-, alanine- and lysine-p-nitroanilide to a similar degree. The most efficient inhibitor was 1,10-phenanthroline. Almost all divalent cations tested inhibited the enzyme activity, while Co2+ stimulated LAP activity by over 100%. The purified LAP had a molecular weight of 90 kDa with an isoelectric point of 5.45. Sodium dodecylsulfate-polyacrylamide gel electrophoresis revealed one band of 48 kDa. (C) 2009 Elsevier Ltd. All rights reserved
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Characterisation of Leucyl Aminopeptidase from Solanum tuberosum tuber
Elsevier Sci Ltd Oxford, 2010Co-Authors: Vujčić Zoran, Dojnov Biljana, Milovanovic Aleksandra, Vujčić Miroslava, Loncar Nikola, Božić NatašaAbstract:Potato juice (a waste product from the starch industry) is a potential source of novel enzymes for food applications. For use in the production and improvement of food protein hydrolysates, commercially available exopeptidases, predominantly Aminopeptidases, are recommended. The present study was performed to explore possible biotechnological interest of Leucyl Aminopeptidase (LAP) activity in the potato tuber. The LAP from potato tuber was purified and characterised. Specific LAP activity was increased 200-fold by purification of the crude extract. The purified enzyme had a pH optimum of 9.0 and temperature optimum of 45 degrees C. LAP hydrolysed leucine-, alanine- and lysine-p-nitroanilide to a similar degree. The most efficient inhibitor was 1,10-phenanthroline. Almost all divalent cations tested inhibited the enzyme activity, while Co2+ stimulated LAP activity by over 100%. The purified LAP had a molecular weight of 90 kDa with an isoelectric point of 5.45. Sodium dodecylsulfate-polyacrylamide gel electrophoresis revealed one band of 48 kDa
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Purification and properties of major midgut Leucyl Aminopeptidase of Morimus funereus (Coleoptera, Cerambycidae) larvae
Elsevier Science Inc New York, 2008Co-Authors: Božić Nataša, Ivanovic Jefisaveta, Nenadovic Vera, Bergstroem Joergen, Larsson Thomas, Vujčić ZoranAbstract:The major Leucyl Aminopeptidase (LAP) from the midgut of Morimus funereus larvae was purified and characterised. Specific LAP activity was increased 292-fold by purification of the crude midgut extract. The purified enzyme had a pH optimum of 7.5 (optimum pH range 7.0-8.5) and preferentially hydrolysed p-nitroanilides containing hydrophobic amino acids in the active site, with the highest V-max /K-M ratio for leucine-p-nitroanilide (LpNA). Among a number of inhibitors tested, the most efficient were 1, 10-phenanthroline having a K-i value of 0.12 mM and cysteine with K-i value of 0.31 mM, while EGTA stimulated LAP activity. Zn2+, Mg2+ and Mn2+ all showed bi-modal effects on LAP activity (activated at low concentrations and inhibited at high concentrations). The purified LAP (after gel filtration on Superose 6 column) had molecular mass of 400 kDa with an isoelectric point of 6.2. Sodium dodecylsulphate-polyacrylamide gel electrophoresis (SDS-PAGE) revealed one band of 67 kDa, suggesting that the enzyme is a hexamer. Six peptide sequences from protein band were obtained using ESI/MS-MS analysis. Comparison of the obtained peptide sequences with the EMBL-EBI sequence analysis toolbox and the BLASTP database showed a high degree of identity with other insect Aminopeptidases. (C) 2007 Elsevier Inc. All rights reserved
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Cytosolic Leucyl Aminopeptidase from the midgut of Morimus funereus larvae
Универзитет у Београду Хемијски факултет, 2007Co-Authors: Božić NatašaAbstract:Cilјеvi оvоg rаdа su bili: rаzviјаnjе visоkооsеtlјivоg i kvаntitаtivnоg zimоgrаmskоg еsеја zаlеucil-аminоpеptidаzе (LАP) i dеtеkciјаizоfоrmi nа оsnоvu zimоgrаmа u sirоvоm еktrаktusrеdnjеg crеvа lаrvi М. funereus; izоlоvаnjеglаvnе izоfоrmе LАP srеdnjеg crеvа lаrvi М.funereus dо hоmоgеnоsti i mоlеkulskа i еnzimskаkаrаktеrizаciја.Оpisаn је оpšti mеtоd zа dеtеkciјu lеucil-аminоpеptidаznе аktivnоsti nаkоn nаtivnеpоliаkrilаmidnе gеl еlеktrоfоrеzе in situ. Меtоdје zаsnivаn nа diаzоtоvаnju p-nitrоаnilinа,оslоbоđеnоg u gеlu dејstvоm lеucil-аminоpеptidаzе nа lеucin-p-nitrоаnilid, kојi јеzаtim kuplоvаn sа hrоmоgеnоm, 1-nаftilаminоm,dо pојаvе ružičаstе аzо-bоје nа mеstu еnzimskеаktivnоsti. Nаđеnо је dа је mеtоd rеprоduktivаn sаkоеficiјеntоm vаriјаciје mаnjim оd 15% zа 32-struki оpsеg, dоk је оbојеnа pоvršinа nа mеstuеnzimskе аktivnоsti u linеаrnој zаvisnоsti оdеnzimskе аktivnоsti.Glаvnа izоfоrmа lеucil-аminоpеptidаzе srеdnjеgcrеvа lаrvi М. funereus је prеčišćеnа dоhоmоgеnоsti i оkаrаktеrisаnа. Spеcifičnааktivnоst LАP је pоrаslа 292 putа prеčišćаvаnjеmsirоvоg еkstrаktа srеdnjеg crеvа. Prеčišćеnеnzim imа pH оptimum 7,5 i prеfеrеnciјаlnоhidrоlizuје p-nitrоаnilidе sа hidrоfоbnimаminоkisеlinаmа, sа nајvеćim Vmax/Km zа lеucin-p-nitrоаnilid. Nајеfikаsniјi inhibitоri su 1,10-fеnаntrоlin sа Ki vrеdnоšću оd 0,12 mМ icistеin sа Ki оd 0,31 mМ. ЕGТА је stimulisаоаktivnоst LАP. Zn2+, Мg2+ i Мn2+ su pоkаzаli bi-mоdаlni еfеkаt nа аktivnоst LАP. PrеčišćеniLАP (nаkоn gеl-filtrаciје nа Superose 6 kоlоni)је imао mоlеkulsku mаsu оd 400 kDа iizоеlеktričnu tаčku 6,2. SDS-PAGE-оm је pоkаzаnајеdnа prоtеinskа trаkа nа 67 kDа, štо ukаzuје dа јееnzim hеksаmеr. Šеst pеptidnih sеkvеnciја јеdоbiјеnо iz prоtеinskе trаkе ЕSI/МS-МS аnаlizоm.Pоrеđеnjеm dоbiјеnih pеptidnih sеkvеnciја ЕМBL-ЕBI prоgrаmоm zа аnаlizu sеkvеnciја sаsеkvеnciјаmа iz BLASTP bаzе pоdаtаkа pоkаzаn јеvisоk stеpеn idеntičnоsti sа drugimаminоpеptidаzаmа insеkаtа.The aims of this work were: development of sensitiveand quantitative zymogram assay for LeucylAminopeptidases (LAP) and detection of LAP isoformsin the crude midgut extract of M. funereus larvae;purification of major LAP from the midgut of M.funereus larvae to homogeneity and its molecular andenzymatic characterization.A general method for detecting Leucyl Aminopeptidaseactivity after native polyacrylamide gel electrophoresisin situ is described. The method is based ondiazotization of p-nitroaniline, liberated in thepolyacrylamide gel by Leucyl Aminopeptidase action onleucine-p-nitroanilide and subsequent coupling with achromogen, 1-naphthylamine, until a pink azo dyeproduct at the position of enzyme activity is obtained.This method was found to be reproducible with thecoefficient of variation below 15% for a 32-fold range,while the colored area of enzyme activity was in lineardependence to enzyme activity.The major Leucyl Aminopeptidase from the midgut of M.funereus larvae was purified and characterised.Specific LAP activity was increased 292-fold bypurification of the crude midgut extract. The purifiedenzyme had a pH optimum of 7,5 and preferentiallyhydrolysed p-nitroanilides containing hydrophobicamino acids in the active site, with the highest Vmax/KMratio for leucine-p-nitroanilide. The most efficientinhibitors were 1,10-phenanthroline having a Ki valueof 0,12 mM and cysteine with Ki value of 0,31 mM.EGTA stimulated LAP activity. Zn2+, Mg2+ and Mn2+ allshowed bi-modal effects on LAP activity. The purifiedLAP (after gel-filtration on Superose 6 column) hadmolecular mass of 400 kDa with an isoelectric point of6,2. SDS-PAGE revealed one band of 67 kDa,suggesting that the enzyme is a hexamer. Six peptidesequences from protein band were obtained usingESI/MS-MS analysis. Comparison of the obtainedpeptide sequences with the EMBL-EBI sequenceanalysis toolbox and the BLASTP database showed ahigh degree of identity with other insectAminopeptidases
Mushtaq A. Saleem - One of the best experts on this subject based on the ideXlab platform.
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In vivo effects of lambda-cyhalothrin on proteases of various body compartments of Periplaneta americana adults.
Pakistan Journal of Zoology, 2010Co-Authors: Mushtaq A. Saleem, Richard M. Wilkins, Abdul Rauf ShakooriAbstract:The overall activity and body comportment distribution of a wide range of cytoplasmic and lysosomal proteolytic enzymes was determined in the adults of American cockroach, Periplaneta americana. Individual proteolytic enzymes showed different overall and relative levels of activity in head, thorax, abdomen, leg, internal leg muscle and gut. The gut showed highest activities of alanyl Aminopeptidase, dipeptidyl Aminopeptidase (DAP) I, II and IV, and cathepsin B, L and H while head showed highest activities of arginyl, Leucyl and tripeptidyl Aminopeptidase. In vivo effects of lambda-cyhalothrin on proteases of various body compartments showed that almost all proteases in the gut, except cathepsin B, showed considerably elevated levels ranging from 19% in the case of proline endopeptidase and 429% in the case of DAP I. All DAPs manifested highest levels i.e. DAP I by 429%, DAP II by 355% and DAP IV by 178%, followed by cathepsin L (246%), cathepsin D (123%), tripeptidyl Aminopeptidase (109%), arginyl Aminopeptidase (78%), alanyl Aminopeptidase (72%), Leucyl Aminopeptidase (49%), cathepsin H (37%) and proline endopeptidase (19%). Proteases of all other body compartments exhibited mixed responses, although in head and thorax, all cytoplasmic proteases were increased while almost all lysosomal proteases were decreased.
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In vivo effects of cyfluthrin on proteolytic enzyme activities of malathion-resistant and susceptible strains of Tribolium castaneum
Pakistan Journal of Zoology, 2004Co-Authors: Mushtaq A. Saleem, David Mantle, Richard M. Wilkins, Abdul Rauf ShakooriAbstract:To elucidate whether insecticide toxicity in insects involves insecticide - induced abnormalities of the intracellular protein catabolic process, we have determined the in vivo effect of a synthetic pyrethroid, cyfluthrin on the activities of representative protein catabolising cytoplasmic and lysosomal proteases (responsible for the various stages of the protein degradation cascade and essential for normal cell functioning) in resistant and susceptible strains of Tribolium castaneum. Effect of cyfluthrin was determined at LC 50 after 48 hour of treatment both in the live and dead adult beetles and compared with controls. In treated live beetles, cyfluthrin decreased alanyl Aminopeptidase (18%), arginyl Aminopeptidase (6%), Leucyl Aminopeptidase (22%), tripeptidyl Aminopeptidase (12%), proline endopeptidase (43%) and dipeptidyl Aminopeptidase I (23%) and increased cathepsin H (42%), while in dead beetles, almost all cytoplasmic proteases as well as cathepsin H manifested further decreasing trend. On the other hand in treated resistant strain live beetles, all lysosomal proteases and Leucyl Aminopeptidase were considerably decreased ranging from 19% to 58% of control activity and this decreasing trend was further intensified in treated dead beetles, which ranged from 30% to 96%. We conclude that the effect of cyfluthrin on proteolytic enzyme activities induced inhibition of proteases and could be important in the development of insecticide resistance in T. castaneum.
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Effect of starvation on proteases in insecticide-resistant and susceptible strains of Musca domestica
Pakistan Journal of Zoology, 2003Co-Authors: Mushtaq A. Saleem, David Mantle, Richard M. Wilkins, Sohail Ahmad, Abdul Rauf ShakooriAbstract:To determine the possible involvement of the process of intracellular protein catabolism in the development of insecticide resistance, we have compared the activities of a comprehensive range of cytoplasmic and lysosomal proteolytic enzymes in 48 hours starved and un-starved resistant and susceptible strains of Musca domestica. Compared to un-starved, the homogenates of 48 hours starved house flies of both resistant and susceptible strains showed elevated levels of most of the proteases tested in this study. Thus in susceptible strain, Leucyl Aminopeptidase, proline endopeptidase and cathepsin L showed elevated levels in starved than un-starved by 42%, 28% and 23%, respectively, while the remaining proteases exhibited minor fluctuations. On the other hand, in resistant strain, Leucyl Aminopeptidase (58%), proline endopeptidase (41%) and cathepsin L (38%) showed elevated levels whereas the remaining proteases exhibited minor deviations. The results suggested that higher induction of proteases in starved resistant than susceptible strain may be important in the development of insecticide resistance in M. domestica.
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Effect of starvation on proteases in insecticide-resistant and susceptible strains of Tribolium castaneum
Pakistan Journal of Zoology, 2003Co-Authors: Mushtaq A. Saleem, David Mantle, Richard M. Wilkins, Abdul Rauf ShakooriAbstract:To elucidate the possible involvement of the process of intracellular protein catabolism in the development of insecticide resistance, we have compared the activities of a comprehensive range of cytoplasmic and lysosomal proteolytic enzymes in 48 hours starved and un-starved resistant (CTC-12) and susceptible (FSS-II) strains of red flour beetle, Tribolium castaneum (Herbst.) (Coleoptera: Tenebrionidae) under laboratory conditions. Compared to un-starved, the homogenates of 48 hours starved adult beetles of both resistant and susceptible strains showed elevated levels of all proteases tested in this study. In starved resistant beetles, all proteases except two were considerably increased which included alanyl Aminopeptidase (41%), arginyl Aminopeptidase (24%), Leucyl Aminopeptidase (154%), dipeptidyl Aminopeptidase IV (27%), tripeptidyl Aminopeptidase (26%), proline endopeptidase (316%), dipeptidyl Aminopeptidase I (153%), dipeptidyl Aminopeptidase II (50%), cathepsin B (22%) and cathepsin L (48%). On the other hand, in starved susceptible beetles, only two proteases viz Leucyl Aminopeptidase (34%) and proline endopeptidase (66%) were increased, while the remaing showed less than 20% elevation. The results suggested that more number and higher induction of proteases in starved resistant than susceptible strain beetles may be important in the development of insecticide-resistance in T. caztaneum.
Richard M. Wilkins - One of the best experts on this subject based on the ideXlab platform.
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In vivo effects of lambda-cyhalothrin on proteases of various body compartments of Periplaneta americana adults.
Pakistan Journal of Zoology, 2010Co-Authors: Mushtaq A. Saleem, Richard M. Wilkins, Abdul Rauf ShakooriAbstract:The overall activity and body comportment distribution of a wide range of cytoplasmic and lysosomal proteolytic enzymes was determined in the adults of American cockroach, Periplaneta americana. Individual proteolytic enzymes showed different overall and relative levels of activity in head, thorax, abdomen, leg, internal leg muscle and gut. The gut showed highest activities of alanyl Aminopeptidase, dipeptidyl Aminopeptidase (DAP) I, II and IV, and cathepsin B, L and H while head showed highest activities of arginyl, Leucyl and tripeptidyl Aminopeptidase. In vivo effects of lambda-cyhalothrin on proteases of various body compartments showed that almost all proteases in the gut, except cathepsin B, showed considerably elevated levels ranging from 19% in the case of proline endopeptidase and 429% in the case of DAP I. All DAPs manifested highest levels i.e. DAP I by 429%, DAP II by 355% and DAP IV by 178%, followed by cathepsin L (246%), cathepsin D (123%), tripeptidyl Aminopeptidase (109%), arginyl Aminopeptidase (78%), alanyl Aminopeptidase (72%), Leucyl Aminopeptidase (49%), cathepsin H (37%) and proline endopeptidase (19%). Proteases of all other body compartments exhibited mixed responses, although in head and thorax, all cytoplasmic proteases were increased while almost all lysosomal proteases were decreased.
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In vivo effects of cyfluthrin on proteolytic enzyme activities of malathion-resistant and susceptible strains of Tribolium castaneum
Pakistan Journal of Zoology, 2004Co-Authors: Mushtaq A. Saleem, David Mantle, Richard M. Wilkins, Abdul Rauf ShakooriAbstract:To elucidate whether insecticide toxicity in insects involves insecticide - induced abnormalities of the intracellular protein catabolic process, we have determined the in vivo effect of a synthetic pyrethroid, cyfluthrin on the activities of representative protein catabolising cytoplasmic and lysosomal proteases (responsible for the various stages of the protein degradation cascade and essential for normal cell functioning) in resistant and susceptible strains of Tribolium castaneum. Effect of cyfluthrin was determined at LC 50 after 48 hour of treatment both in the live and dead adult beetles and compared with controls. In treated live beetles, cyfluthrin decreased alanyl Aminopeptidase (18%), arginyl Aminopeptidase (6%), Leucyl Aminopeptidase (22%), tripeptidyl Aminopeptidase (12%), proline endopeptidase (43%) and dipeptidyl Aminopeptidase I (23%) and increased cathepsin H (42%), while in dead beetles, almost all cytoplasmic proteases as well as cathepsin H manifested further decreasing trend. On the other hand in treated resistant strain live beetles, all lysosomal proteases and Leucyl Aminopeptidase were considerably decreased ranging from 19% to 58% of control activity and this decreasing trend was further intensified in treated dead beetles, which ranged from 30% to 96%. We conclude that the effect of cyfluthrin on proteolytic enzyme activities induced inhibition of proteases and could be important in the development of insecticide resistance in T. castaneum.
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Effect of starvation on proteases in insecticide-resistant and susceptible strains of Musca domestica
Pakistan Journal of Zoology, 2003Co-Authors: Mushtaq A. Saleem, David Mantle, Richard M. Wilkins, Sohail Ahmad, Abdul Rauf ShakooriAbstract:To determine the possible involvement of the process of intracellular protein catabolism in the development of insecticide resistance, we have compared the activities of a comprehensive range of cytoplasmic and lysosomal proteolytic enzymes in 48 hours starved and un-starved resistant and susceptible strains of Musca domestica. Compared to un-starved, the homogenates of 48 hours starved house flies of both resistant and susceptible strains showed elevated levels of most of the proteases tested in this study. Thus in susceptible strain, Leucyl Aminopeptidase, proline endopeptidase and cathepsin L showed elevated levels in starved than un-starved by 42%, 28% and 23%, respectively, while the remaining proteases exhibited minor fluctuations. On the other hand, in resistant strain, Leucyl Aminopeptidase (58%), proline endopeptidase (41%) and cathepsin L (38%) showed elevated levels whereas the remaining proteases exhibited minor deviations. The results suggested that higher induction of proteases in starved resistant than susceptible strain may be important in the development of insecticide resistance in M. domestica.
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Effect of starvation on proteases in insecticide-resistant and susceptible strains of Tribolium castaneum
Pakistan Journal of Zoology, 2003Co-Authors: Mushtaq A. Saleem, David Mantle, Richard M. Wilkins, Abdul Rauf ShakooriAbstract:To elucidate the possible involvement of the process of intracellular protein catabolism in the development of insecticide resistance, we have compared the activities of a comprehensive range of cytoplasmic and lysosomal proteolytic enzymes in 48 hours starved and un-starved resistant (CTC-12) and susceptible (FSS-II) strains of red flour beetle, Tribolium castaneum (Herbst.) (Coleoptera: Tenebrionidae) under laboratory conditions. Compared to un-starved, the homogenates of 48 hours starved adult beetles of both resistant and susceptible strains showed elevated levels of all proteases tested in this study. In starved resistant beetles, all proteases except two were considerably increased which included alanyl Aminopeptidase (41%), arginyl Aminopeptidase (24%), Leucyl Aminopeptidase (154%), dipeptidyl Aminopeptidase IV (27%), tripeptidyl Aminopeptidase (26%), proline endopeptidase (316%), dipeptidyl Aminopeptidase I (153%), dipeptidyl Aminopeptidase II (50%), cathepsin B (22%) and cathepsin L (48%). On the other hand, in starved susceptible beetles, only two proteases viz Leucyl Aminopeptidase (34%) and proline endopeptidase (66%) were increased, while the remaing showed less than 20% elevation. The results suggested that more number and higher induction of proteases in starved resistant than susceptible strain beetles may be important in the development of insecticide-resistance in T. caztaneum.