Affinity purification and binding analysis of the hemolymph juvenile hormone binding protein from Manduca sexta

Park, Yong Chul; Tesch, Michael J.; Toong, Yock Chai; Goodman, Walter G.

doi:10.1021/bi00082a011

Cited by 31 publications

(17 citation statements)

References 37 publications

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“…As a positive control, the same amount of tritiated JH was incubated with 250l hemolymph from Manduca sexta fourth instar larvae, which contains a 34-36kDa JH binding protein, JHBP (Goodman et al, 1978;Park et al, 1993;Orth et al, 2003). After incubation, samples of BSA and M. sexta hemolymph were separated in duplicate on 10% native gels at 180V.…”

Section: Jh Assays Juvenile Hormone Binding Assaymentioning

confidence: 99%

Insulin-like peptide genes in honey bee fat body respond differently to manipulation of social behavioral physiology

Nilsen

Ihle

Frederick

et al. 2011

Journal of Experimental Biology

107

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SUMMARY Nutrient sensitive insulin-like peptides (ILPs) have profound effects on invertebrate metabolism, nutrient storage, fertility and aging. Many insects transcribe ILPs in specialized neurosecretory cells at changing levels correlated with life history. However, the major site of insect metabolism and nutrient storage is not the brain, but rather the fat body, where functions of ILP expression are rarely studied and poorly understood. Fat body is analogous to mammalian liver and adipose tissue, with nutrient stores that often correlate with behavior. We used the honey bee (Apis mellifera), an insect with complex behavior, to test whether ILP genes in fat body respond to experimentally induced changes of behavioral physiology. Honey bee fat body influences endocrine state and behavior by secreting the yolk protein precursor vitellogenin (Vg), which suppresses lipophilic juvenile hormone and social foraging behavior. In a two-factorial experiment, we used RNA interference (RNAi)-mediated vg gene knockdown and amino acid nutrient enrichment of hemolymph (blood) to perturb this regulatory module. We document factor-specific changes in fat body ilp1 and ilp2 mRNA, the bee's ILP-encoding genes, and confirm that our protocol affects social behavior. We show that ilp1 and ilp2 are regulated independently and differently and diverge in their specific expression-localization between fat body oenocyte and trophocyte cells. Insect ilp functions may be better understood by broadening research to account for expression in fat body and not only brain.

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Section: Jh Assays Juvenile Hormone Binding Assaymentioning

confidence: 99%

Insulin-like peptide genes in honey bee fat body respond differently to manipulation of social behavioral physiology

Nilsen

Ihle

Frederick

et al. 2011

Journal of Experimental Biology

107

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“…The molecular identification of the to gene has revealed that it shares structural homology with JHBPs (Sarov-Blat et al, 2000). JH, synthesized in the corpus allatum, circulates in haemolymph bound to JHBPs, which serve as carriers for this hydrophobic hormone (Park et al, 1993). Further investigation needs to be done to demonstrate that TO could be a carrier for JH.…”

Section: Why Do To 1 Flies Die Faster Than Wt During Starvation?mentioning

confidence: 99%

Regulation of feeding behaviour and locomotor activity bytakeoutinDrosophila

Meunier

Belgacem

Martin

2007

Journal of Experimental Biology

140

143

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SUMMARY The hormonal regulation of feeding behaviour is well known in vertebrates,whereas it remains poorly understood in insects. Here, we report that the takeout gene is an essential component of nutritional homeostasis in Drosophila. takeout encodes a putative juvenile hormone (JH)binding protein and has been described as a link between circadian rhythm and feeding behaviour. However, the physiological role of takeout and its putative link to JH remain unknown. In this study, we show that takeout (to1) flies failed to adapt their food intake according to food availability and that most defects could be genetically rescued. When food is abundant, to1 are hyperphagic, yielding to hypertrophy of the fat body. When food reappears after a starvation period, to1 flies do not increase their food intake as much as wild-type flies. This defect in food intake regulation is partly based on the action of Takeout on taste neurons, because the sensitivity of to1 gustatory neurons to sugars does not increase after starvation, as in wild-type neurons. This lack of regulation is also evident at the locomotor activity, which normally increases during starvation, a behaviour related to food foraging. In addition, to1 flies lack sexual dimorphism of locomotor activity,which has previously been linked to the JH circulating level. Moreover,application of the JH analog methoprene rescues the phenotype. These results suggest that takeout plays a central role as a feeding regulator and may act by modulating the circulating JH level.

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“…We also assayed JH binding by the DEAE-cellulose filter binding assay (Peterson, 1985) and equilibrium dialysis (Park et al, 1993).…”

Section: Juvenile Hormone Binding Assaysmentioning

confidence: 99%

Purification and reassessment of ligand binding by the recombinant, putative juvenile hormone receptor of the tobacco hornworm,Manduca sexta

Charles

Wojtasek

Lentz

et al. 1996

Arch. Insect Biochem. Physiol.

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The 29 kDa protein from the larval epidermis of the tobacco hornworm, Manduca sexta, that specifically bound photoaffinity analogs of JH I and JH II was produced by a recombinant baculovirus (rJP29). The higher of the two molecular weight forms made corresponded to a protein that could be formed by read‐through of the TGA termination codon to the following TAA. The previously reported, apparent high affinity binding of [methyl‐3H]‐JH I by rJP29 as measured by the dextran‐coated charcoal (DCC) assay [Palli et al., Proc Natl Acad Sci USA 91:6191–6195 (1994)] was found to be artifactual due to endogenous cellular esterases that co‐purified with rJP29 through both DEAE cellulose and MonoQ chromatography. These esterases converted the 10–20 nM labelled JH to JH I acid and [3H]‐methanol during the 1 h incubation at room temperature. Additionally, DEAE fractions containing rJP29 or from wild‐type virus‐infected cells were found to bind nonspecifically high amounts of 12, 13‐3H]‐JH I acid in the DCC assay. Neither rJP29 nor the cellular esterases had JH esterase activity when assayed on a series of thioether surrogate substrates. When separated from these contaminating esterases either by hydroxylapatite or affinity chromatography, rJP29 showed little or no detectable binding of [12,13‐3H]‐JH I. Yet purified rJP29 bound EBDA, the photoaffinity analog of JH I; and this binding was prevented by retinol, JH I, methyl farnesoate, methoprene, and xanthophyll, but not by farnesol and 20‐hydroxyecdysone. Therefore, JP29 is not a high affinity JH receptor. © 1996 Wiley‐Liss, Inc.

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Affinity purification and binding analysis of the hemolymph juvenile hormone binding protein from Manduca sexta

Cited by 31 publications

References 37 publications

Insulin-like peptide genes in honey bee fat body respond differently to manipulation of social behavioral physiology

Insulin-like peptide genes in honey bee fat body respond differently to manipulation of social behavioral physiology

Regulation of feeding behaviour and locomotor activity bytakeoutinDrosophila

Purification and reassessment of ligand binding by the recombinant, putative juvenile hormone receptor of the tobacco hornworm,Manduca sexta

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