Characterization of azadirachtin binding to Sf9 nuclei in vitro

Nisbet, Alasdair J.; Mordue, A. J.; Grossman, Robert B.; Jennens, Lyn; Ley, Steven V.; Mordue, W.

doi:10.1002/(sici)1520-6327(1997)34:4<461::aid-arch5>3.0.co;2-v

Cited by 16 publications

(10 citation statements)

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“…Inhibition of cell proliferation in the presence of azadirachtin has been observed for several insect cell lines (Rembold and Annadurai, 1993;Nisbet et al, 1997;Salehzadeh et al, 2002). In this study, the same effects of inhibition of cell proliferation were observed for Drosophila cell line, Kc167, when incubated with various doses of azadirachtin for 48 h, resulting in an ED 50 value of 1.7 × 10 -7 M, which is similar to that obtained by others (Rembold and Annadurai, 1993;Nisbet et al, 1997;Salehzadeh et al, 2002).…”

Section: Discussionsupporting

confidence: 90%

“…Azadirachtin binding was not disrupted by agents that cause depolymerisation of the microtubules but was disrupted by a variety of detergents, proteinases, and heat-treatment, suggesting that the binding site may be part of a group of proteins associated with the cytoskeleton but not necessarily cytoskeletal in origin (Mordue (Luntz) et al, 1999). This knowledge that azadirachtin binds preferentially to the nuclear extract of Sf9 cells (Nisbet et al, 1997) and that the "receptor" is possibly a protein complex, the function of which is easily disrupted by detergents, forms the basis for further identification and characterization of the specific binding site for azadirachtin in insect cell lines in this study. The cell line of choice was the Drosophila melanogaster cell line Kc167 because of the relative ease with which any protein or protein complex identified could be screened for and potentially identified using the Drosophila flybase (http://flybase.bio.…”

Section: Introductionmentioning

confidence: 89%

“…In Sf9 cell (derived from pupal ovarian tissue of the lepidopteran, Spodoptera frugiperda) subcellular fractions, [22, H 2 ] dihydroazadirachtin predominantly bound to components of the nuclear fraction suggesting that it contains the azadirachtinbinding component(s) (Rembold and Annadurai, 1993;Nisbet et al, 1997;Salehzadeh et al, 2003). The binding site was further characterised in terms of affinity and specificity by Nisbet et al (1995Nisbet et al ( , 1996Nisbet et al ( , 1997 and Mordue (Luntz) et al (1999). Azadirachtin binding was not disrupted by agents that cause depolymerisation of the microtubules but was disrupted by a variety of detergents, proteinases, and heat-treatment, suggesting that the binding site may be part of a group of proteins associated with the cytoskeleton but not necessarily cytoskeletal in origin (Mordue (Luntz) et al, 1999).…”

Section: Introductionmentioning

confidence: 99%

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Identification of a putative azadirachtin‐binding complex from Drosophila Kc167 cells

Robertson

Ni²,

Dhadialla³

et al. 2007

Arch Insect Biochem Physiol

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Cell-proliferation in Drosophila Kc167 cells was inhibited by 50% when cell cultures contained 1.7 x 10(-7) M azadirachtin for 48 h (a tertranortriterpenoid from the neem tree Azadirachta indica). Drosophila Kc167 cells exhibited direct nuclear damage within 6-h exposure to azadirachtin (5 x 10(-7) M and above) or within 24 h when lower concentrations were used (1 x 10(-9) M). Fractionation of an extract of Drosophila Kc167 cells combined with ligand overlay technique resulted in the identification of a putative azadirachtin binding complex. Identification of the members of this complex by Peptide Mass Fingerprinting (PMF) and N-terminal sequencing identified heat shock protein 60 (hsp60) as one of its components.

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Section: Discussionsupporting

confidence: 90%

Section: Introductionmentioning

confidence: 89%

Section: Introductionmentioning

confidence: 99%

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Identification of a putative azadirachtin‐binding complex from Drosophila Kc167 cells

Robertson

Ni²,

Dhadialla³

et al. 2007

Arch Insect Biochem Physiol

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“…Their efficacy is directly related to azadirachtin content however many of the other compound also have biological activity and add to its effects. whereas pure azadirachtin has been shown to be effective in the field (Mordue et al 1997) the natural mixtures of azadirachtins in neem insecticides may usefully mitigate against the development of resistance compared to azadirachtin alone (Feng & Isman 1995).…”

Section: Chemistrymentioning

confidence: 99%

“…Sub-cellular fractionation of Sf9 cells (captured insect cells derived from S. frugiperda) incubated with [22, H 2 ] dihydroazadirachtin during logarithmic growth phase revealed high affinity specific binding to the nuclear fraction of the cells (Nisbet et al 1997). A comparison of binding of tritiated dihydroazadirachtin to these two insect tissues shows specific, time-dependent, saturable high affinity binding in both tissues, with many similarities in binding characteristics (Table 3).…”

Section: Understanding the Effects Of Azadirachtin On Insectsmentioning

confidence: 99%

Azadirachtin from the neem tree Azadirachta indica: its action against insects

Mordue

Nisbet

2000

An. Soc. Entomol. Bras.

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Azadiractina do Nim, Azadirachta indica: sua Ação Contra Insetos RESUMO -A árvore do nim há muito tempo é reconhecida por suas propriedades singulares de ação contra insetos e benefício à saúde humana. É plantada na maior parte das áreas tropicais e subtropicais do mundo para sombra, reflorestamento e produção de matéria prima para inseticidas naturais e medicamentos. A azadiractina, complexo tetranortriterpenóide limonóide das sementes é o principal composto responsável pelos efeitos tóxicos aos insetos. Seis conferências internacionais sobre nim e vasta literatura científica relatam esses aspectos. Este artigo revê as propriedades da azadiractina no comportamento e na fisiologia de insetos, incluindo os efeitos na reprodução, redução da alimentação tanto direto quanto a chamada "secundária", redução do crescimento, aumento da mortalidade e ocorrência de ecdises anormais e tardias. Os efeitos fisiológicos são aqui categorizados de duas maneiras: efeitos diretos sobre as células e os tecidos e efeitos indiretos exercidos via o sistema endócrino. O artigo também descreve o trabalho feito até o momento visando identificar o modo de açao da azadiractina em nível celular e seus efeitos diferentes entre filos animais e sobre organismos não nocivos, o que indica seu sucesso potencial como inseticida seguro. PALAVRAS-CHAVE: Insecta, inseticida botânico, fisiologia de insetos.ABSTRACT -The neem tree has long been recognized for its unique properties both against insects and in improving human health. It is grown in most tropical and sub-tropical areas of the world for shade, reforestation and for the production of row material for natural insecticides and medicines. Azadirachtin, a complex tetranortriterpenoid limonoid from the neem seeds, is the main component responsible for the toxic effects in insects. Six international conferences on neem and a vast scientific literature report both the antifeedant and physiological effects of neem. This article reviews the behavioral and physiological properties of azadirachtin, including effects on insect reproduction, direct and "secondary" antifeedancy, and the physiological effects measured as growth reduction, increased mortality and abnormal and delayed moults. These effects are here categorized in two ways: direct effects on cells and tissues and indirect effects exerted via the endocrine system. It also describes the work carried out to date to identify the mode of action of azadirachtin at the cellular level. The differential effects between animal phyla and over non-target organisms are discussed and point to its potential success as a safe insecticide.

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Actions of azadirachtin, a plant allelochemical, against insects

et al. 1998

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Investigations of the antifeedant mode of action of azadirachtin and four synthetic analogues, 22,23-dihydroazadirachtin, 3-tigloylazadirachtol, 11methoxydihydroazadirachtin and 22,23-bromoethoxydihydroazadirachtin have revealed that both polyphagous and oligophagous insects are behaviourally responsive to azadirachtin, with the most responsive species being able to di †erentiate extremely small changes in the parent molecule. In Lepidoptera the antifeedant response is correlated also with increased neural activity of the chemoreceptors. When locusts are treated on crop plants, the antifeedant and physiological actions of azadirachtin and analogues work in concert and result in feeding deterrence, growth and moulting aberrations and mortality with the same order of potency as for antifeedancy. SpeciÐc binding studies using [3H]dihydroazadirachtin carried out on locust testes and Spodoptera Sf9 cells have shown that the competitive binding of the di †erent analogues of azadirachtin to these binding sites occurs in a similar order of potency to that found with antifeedant and IGR bioassays. This suggests a causal link between speciÐc binding to membrane proteins and the ability of the molecule to exert biological e †ects.1998 Society of Chemical Industry ( Pestic. Sci., 54, 277È284 (1998)

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Characterization of azadirachtin binding to Sf9 nuclei in vitro

Cited by 16 publications

References 18 publications

Identification of a putative azadirachtin‐binding complex from Drosophila Kc167 cells

Identification of a putative azadirachtin‐binding complex from Drosophila Kc167 cells

Azadirachtin from the neem tree Azadirachta indica: its action against insects

Actions of azadirachtin, a plant allelochemical, against insects

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