A novel chitin-binding protein identified from the peritrophic membrane of the cabbage looper, Trichoplusia ni

Guo, Wei; Li, Guoxun; Pang, Yi; Wang, Ping

doi:10.1016/j.ibmb.2005.06.003

Cited by 86 publications

(111 citation statements)

References 35 publications

(56 reference statements)

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“…Whether this activity is due to the physical interaction of HaCDA5a disrupting the PM, to its hypothetical deacetylation activity, or to any other enzymatic activity is something yet to be determined. We were unable to demonstrate deacetylase activity of HaCDA5a by measuring chitin deacetylation by native gel electrophoresis (41; data not shown), confirming previous results with TnPM-P42 CDA from T. ni (13). In contrast, Toprak et al (40), using a similar methodology, have shown deacetylase activity of a 26-kDa form of the HaCDA5a ortholog from M. configurata (MacoCDA) recombinantly expressed in E. coli.…”

Section: Discussionsupporting

confidence: 86%

“…Chitin binding activity assay. Due to the facts that the HaCDA5a homolog in Trichoplusia ni was found in the PM (13) and that the PM consists of proteins and chitin, the ability of HaCDA5a to bind chitin was checked. Recombinant HaCDA5a expressed in Sf21 cells and secreted into the cell culture medium exhibited a strong binding affinity for regenerated chitin.…”

Section: Resultsmentioning

confidence: 99%

“…Also, for T. ni, an HaCDA5a ortholog was identified in the larval midgut (13). The availability of the whole B. mori genome enabled the identification of eight chitin deacetylase homologs in this species.…”

Section: Discussionmentioning

confidence: 99%

“…HaCDA5a, which was demonstrated to bind chitin, contains 15 cysteines along its sequence, but their distribution does not resemble the C6 motif. It was proposed that the CDA domain may serve as a chitin binding domain (13). Alternative non-cysteine interactions with chitin have been described for arthropods (32).…”

Section: Discussionmentioning

confidence: 99%

“…The chitin binding activity of HaCDA5a was checked according to a previously described method (13). Chitin was regenerated as follows.…”

Section: Semiquantitative and Real-time Quantitative Rt-pcrmentioning

confidence: 99%

See 4 more Smart Citations

Downregulation of a Chitin Deacetylase-Like Protein in Response to Baculovirus Infection and Its Application for Improving Baculovirus Infectivity

Jakubowska

Caccia

Gordon

et al. 2010

J Virol

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Several expressed sequence tags (ESTs) with homology to chitin deacetylase-like protein (CDA) were selected from a group of Helicoverpa armigera genes whose expression changed after infection with H. armigera single nucleopolyhedrovirus (HearNPV). Some of these ESTs coded for a midgut protein containing a chitin deacetylase domain (CDAD). The expressed protein, HaCDA5a, did not show chitin deacetylase activity, but it showed a strong affinity for binding to chitin. Sequence analysis showed the lack of any chitin binding domain, described for all currently known peritrophic membrane (PM) proteins. HaCDA5a has previously been detected in the H. armigera PM. Such localization, together with its downregulation after pathogen infection, led us to hypothesize that this protein might be responsible for the homeostasis of the PM structure and that, by reduction of its expression, the insect may reduce PM permeability, decreasing the entrance of baculovirus. To test this hypothesis, we constructed a recombinant nucleopolyhedrovirus to express HaCDA5a in insect cells and tested its influence on PM permeability as well as the influence of HaCDA5a expression on the performance of the baculovirus. The experiments showed that HaCDA5a increased PM permeability, in a concentration-dependent manner. Bioassays on Spodoptera frugiperda and Spodoptera exigua larvae revealed that NPV expressing HaCDA5a was more infective than its parental virus. However, no difference in virulence was observed when the viruses were injected intrahemocoelically. These findings support the downregulation of a midgut-specific CDA-like protein as a possible mechanism used by H. armigera to reduce susceptibility to baculovirus by decreasing PM permeability.

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Section: Discussionsupporting

confidence: 86%

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

“…The chitin binding activity of HaCDA5a was checked according to a previously described method (13). Chitin was regenerated as follows.…”

Section: Semiquantitative and Real-time Quantitative Rt-pcrmentioning

confidence: 99%

See 3 more Smart Citations

Downregulation of a Chitin Deacetylase-Like Protein in Response to Baculovirus Infection and Its Application for Improving Baculovirus Infectivity

Jakubowska

Caccia

Gordon

et al. 2010

J Virol

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Identification and characterization of two chitin‐binding proteins from the peritrophic membrane of the silkworm, Bombyx mori L.

Yang

Zhou

Malik

et al. 2010

Arch Insect Biochem Physiol

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The peritrophic membrane (PM) is a semi-permeable lining of the insect midgut, broadly analogous to the mucous lining of vertebrate gut. The PM proteins are important achievements for the function of the PM. In this study, two chitin-binding proteins (BmPM-P43 and BmPM-P41) from the PM of the silkworm, Bombyx mori, were identified and cloned. These proteins showed the molecular mass of 43 and 41 kDa, respectively. The deduced amino acid sequences codes for a protein of 381 amino acid residues and 364 amino acid residues, containing 12 and 14 cysteine residues followed by similar domain, both of them have 5 cysteine residues in similar position in the C-terminal. The confirmation of these proteins was performed by western blot analysis of recombinant BmPM-P43 and BmPM-P41. The chitin-binding activity analysis showed that the BmPM-P43 and BmPM-P41 could bind to chitin strongly. It is concluded that BmPM-P43 and BmPM-P41 contains a polysaccharide deacetylase domain instead of peritrophin domain, indicated that these two proteins may belong to a new chitin-binding protein family.

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CONSTRUCTION OF SILKWORM MIDGUT cDNA LIBRARY FOR SCREEN AND SEQUENCE ANALYSIS OF PERITROPHIC MEMBRANE PROTEIN GENES

Zhou

Xue

et al. 2015

Arch Insect Biochem Physiol

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Silkworm is an important economic insect and the model species for Lepidoptera. The midgut of silkworm is an important physiological barrier, as its peritrophic membrane (PM) can resist pathogen invasion. In this study, a silkworm midgut cDNA library was constructed in order to identify silkworm PM genes. The capacity of the initial library was 6.92 × 10(6) pfu/ml, along with a recombination rate of 92.14% and a postamplification titer of 4.10 × 10(9) pfu/ml. Three silkworm PM protein genes were obtained by immunoscreening, two of which were chitin-binding protein (CBP) genes and one of which was a chitin deacetylase (CDA) gene as revealed by sequence analysis. Three genes were named BmCBP02, BmCBP13, and BmCDA17, and their ORF sizes are 678, 1,029, and 645 bp, respectively; all of them contain sequences of chitin-binding domains. Phylogenetic analysis indicated that BmCBP02 has the highest consensus with Mamestra configurata CBP at 61.0%; BmCBP13 has the highest consensus with Loxostege sticticalis PM CBP at 53.35%; BmCDA17 has the highest consensus with Helicoverpa armigera CDA5a at 70.83%. Tissue transcriptional analysis revealed that all three genes were specifically expressed in the midgut, and during the developmental process of fifth-instar silkworms, the transcription of all the genes showed an upward trend. This study laid a foundation for further studies on the functions of silkworm PM genes.

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A novel chitin-binding protein identified from the peritrophic membrane of the cabbage looper, Trichoplusia ni

Cited by 86 publications

References 35 publications

Downregulation of a Chitin Deacetylase-Like Protein in Response to Baculovirus Infection and Its Application for Improving Baculovirus Infectivity

Downregulation of a Chitin Deacetylase-Like Protein in Response to Baculovirus Infection and Its Application for Improving Baculovirus Infectivity

Identification and characterization of two chitin‐binding proteins from the peritrophic membrane of the silkworm, Bombyx mori L.

CONSTRUCTION OF SILKWORM MIDGUT cDNA LIBRARY FOR SCREEN AND SEQUENCE ANALYSIS OF PERITROPHIC MEMBRANE PROTEIN GENES

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