Proteomic analysis of the peritrophic matrix from the gut of the caterpillar, Helicoverpa armigera

Campbell, Peter M.; Cao, Anh T.; Hines, Eric R.; East, Peter D.; Gordon, Karl

doi:10.1016/j.ibmb.2008.07.009

Cited by 103 publications

(115 citation statements)

References 59 publications

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“…Certainly glycosylation of other lepidopteran APNs is demonstrated (Knight et al, 2004;Sangadala et al, 2001), toxin-binding forms of an H. virescens APN differed by about 40 kDa (Oltean et al, 1999), and we observed distinctly reduced apparent molecular weights for the four H. armigera APNs when they were exposed to a deglycosylation treatment that would remove both N-and O-linked glycan (Edge, 2003;Campbell et al, 2008).…”

Section: Discussionmentioning

confidence: 54%

“…The proteins were identified by comparing the mass spectra of tryptic digest peptides with a database of genes expressed in the midgut of H. armigera. The polycalin and four APNs reported here were also recovered from the peritrophic matrix (PM) of H. armigera (Campbell et al, 2008) (Fig. 3).…”

Section: Discussionmentioning

confidence: 62%

“…All the APN sequences predict proteins of about 110 kDa so the observed gel mobilities are consistent with variable additions of glycan. In another study (Campbell et al, 2008) we identified HaAPNs from SDS-PAGE bands after a chemical deglycosylation treatment (Edge, 2003). The APNs migrated in the same order (HaAPN1 and HaAPN2 comigrated, HaAPN3 migrated a little further and HaAPN4 was the furthest) but all migrated a little further than a 97 kDa marker protein.…”

Section: Resultsmentioning

confidence: 96%

“…In H. armigera the observed mobility on SDS-PAGE of the affinity purified polycalin was consistent with the predicted size of the longest, pentacalin sequence, and there were no peptides that were unique matches with either of the shorter cDNA sequences. However, polycalin-derived peptides were recovered from SDS-PAGE of an extract of the peritrophic matrix across a very wide size range (retained in the first millimetre of the gel and gel slices in the range 40-150 kDa, Campbell et al, 2008).…”

Section: Discussionmentioning

confidence: 99%

“…These were also processed by in-gel digestion and LC/MS analysis, this time with an Agilent XCT ion trap mass spectrometer as described elsewhere (Campbell et al, 2001(Campbell et al, , 2008. In the intervening period, a more extensive collection of EST and full-length cDNA sequences from the midgut of H. armigera was developed (below) against which we could compare both the earlier peptide sequence interpretations and the later ion trap MS peptide fragmentation data.…”

Section: Identification Of Protein Bandsmentioning

confidence: 99%

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Diversity of aminopeptidases, derived from four lepidopteran gene duplications, and polycalins expressed in the midgut of Helicoverpa armigera: Identification of proteins binding the δ-endotoxin, Cry1Ac of Bacillus thuringiensis

Angelucci

Barrett-Wilt²,

Hunt

et al. 2008

Insect Biochemistry and Molecular Biology

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Helicoverpa armigera midgut proteins that bind the Bacillus thuringiensis (Bt) δ-endotoxin Cry1Ac were purified by affinity chromatography. SDS-PAGE showed that several proteins were eluted with N-acetylgalactosamine and no further proteins were detected after elution with urea. Tandem mass spectral data for tryptic peptides initially indicated that the proteins resembled aminopeptidases (APNs) from other lepidopterans and cDNA sequences for seven APNs were isolated from H. armigera through a combination of cloning with primers derived from predicted peptide sequences and established EST libraries. Phylogenetic analysis showed lepidopteran APN genes in nine clades of which five were part of a lepidopteran-specific radiation. The Cry1Ac-binding proteins were then identified with four of the seven HaAPN genes. Three of those four APNs are likely orthologs of APNs characterised as Cry1Ac-binding proteins in other lepidopterans. The fourth Cry1Ac-binding APN has orthologs not previously identified as Cry1Ac-binding partners. The HaAPN genes were expressed predominantly in the midgut through larval development. Each showed consistent expression along the length of the midgut but five of the genes were expressed at levels about two orders of magnitude greater than the remaining two. The remaining mass spectral data identified sequences encoding polycalin proteins with multiple lipocalin-like domains. A polycalin has only been previously reported in another lepidopteran, Bombyx mori, but polycalins in both species are now linked with binding of Bt Cry toxins. This is the first report of hybrid, lipocalin-like domains in shorter polycalin sequences that are not present in the longest sequence. We propose that these hybrid domains are generated by alternative splicing of the mRNA.

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Section: Discussionmentioning

confidence: 54%

Section: Discussionmentioning

confidence: 62%

Section: Resultsmentioning

confidence: 96%

Section: Discussionmentioning

confidence: 99%

Section: Identification Of Protein Bandsmentioning

confidence: 99%

See 3 more Smart Citations

Diversity of aminopeptidases, derived from four lepidopteran gene duplications, and polycalins expressed in the midgut of Helicoverpa armigera: Identification of proteins binding the δ-endotoxin, Cry1Ac of Bacillus thuringiensis

Angelucci

Barrett-Wilt²,

Hunt

et al. 2008

Insect Biochemistry and Molecular Biology

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Characterization of the trypsin‐like protease (Ha‐TLP2) constitutively expressed in the integument of the cotton bollworm, Helicoverpa armigera

Liu

Sui

Wang

et al. 2009

Arch Insect Biochem Physiol

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Trypsins belong to the serine endoproteases. They are the most important proteases in insects because of their key roles in food digestion and zymogens activation. But there has been little study of the trypsins in the integuments of insects. In this work, we cloned a trypsin-like protease gene from Helicoverpa armigera and named it trypsin-like protease 2 (Ha-TLP2). Semi-quantitative reverse transcription PCR analysis showed that Ha-TLP2 is constitutively expressed in the integument and can be down-regulated by 20-hydroxyecdysone (20E) and up-regulated by the juvenile hormone (JH) analog methoprene. Immunohistochemistry showed that Ha-TLP2 is located not only in the epidermis, but also in new and old cuticles. Immunoblotting and gelatin-SDS-PAGE revealed that Ha-TLP2 is constitutively expressed with activity in the integument during larval feeding, molting, and metamorphosis. This evidence suggests that Ha-TLP2 is involved in the remodeling of the integument.

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CLONING AND CHARACTERIZATION OF A MIDGUT‐SPECIFIC FATTY ACID BINDING PROTEIN IN Spodoptera litura

Huang

Zhou

Gao

et al. 2012

Arch Insect Biochem Physiol

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A fatty acid binding protein (FABP) gene (Slfabp1) was cloned from the midgut of Spodoptera litura larvae. The gene consists of four exons and three introns and encodes a peptide of 134 amino acid residues with a predicted molecular mass of 14.7 kDa, which was confirmed by in vitro protein expression. Northern blot and Western blot analyses indicated that both of Slfabp1 mRNA and protein were highly and specifically expressed in the midgut during the fifth and sixth instar feeding larval stages. In situ hybridization and immunohistochemistry analyses confirmed the midgut-specific localization of Slfabp1 mRNA and protein. The result of Western blot showed that expression of the protein was downregulated by starvation and upregulated by refeeding in sixth instar larvae. All of the results taken together suggest that the SlFABP1 plays important role(s) in FA uptake and transport in the midgut during the larval feeding stages of the insect.

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Proteomic analysis of the peritrophic matrix from the gut of the caterpillar, Helicoverpa armigera

Cited by 103 publications

References 59 publications

Diversity of aminopeptidases, derived from four lepidopteran gene duplications, and polycalins expressed in the midgut of Helicoverpa armigera: Identification of proteins binding the δ-endotoxin, Cry1Ac of Bacillus thuringiensis

Diversity of aminopeptidases, derived from four lepidopteran gene duplications, and polycalins expressed in the midgut of Helicoverpa armigera: Identification of proteins binding the δ-endotoxin, Cry1Ac of Bacillus thuringiensis

Characterization of the trypsin‐like protease (Ha‐TLP2) constitutively expressed in the integument of the cotton bollworm, Helicoverpa armigera

CLONING AND CHARACTERIZATION OF A MIDGUT‐SPECIFIC FATTY ACID BINDING PROTEIN IN Spodoptera litura

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