Isolation of ferritin and its interaction with BmNPV in the silkworm, Bombyx mori

Fei, Dong-Qiong; Yu, Hai‐Zhong; Xu, Jia-ping; Zhang, Shang-zhi; Wang, Jie; Li, Bing; Yang, Liang; Hu, Pei; Zhao, Kang; Toufeeq, Shahzad

doi:10.1016/j.dci.2018.05.012

Cited by 13 publications

(13 citation statements)

References 45 publications

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“…After iron ion treatment, upregulated expression of ScFerLCH was found in the midgut, hemolymph, and fat body; thus, we inferred that the coexpressed ScFerLCH would also be induced after iron treatment. Similar result were also observed in B. mori (Fei et al, 2018) and many other species (Nichol, Law, & Winzerling, 2002).…”

Section: Discussionsupporting

confidence: 85%

“…The first day of the fifth‐instar larvae was fed 10% ferric chloride (Sangon Biotech; prepared with sterile water) for 24 hr, and then the hemolymph was collected and stored at −20°C for later use. Ferritin protein was extracted from the hemolymph according to a previously published method (Fei et al, 2018; Nichol & Locke, 1989), with some modifications. Briefly, 300 μl hemolymph samples were added to 3.5 ml precooling extraction buffer (0.1% Triton X‐100, 0.5 M Tris‐HCl [pH 6.8], 0.2% ferene disodium salt, and 1 mM phenyl methane sulfonyl fluoride [PMSF]) and homogenized.…”

Section: Methodsmentioning

confidence: 99%

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Identification of FerLCH, isolation of ferritin and functional analysis related to interaction with pathogens in Eri‐silkworm, Samia cynthia ricini

Yang

Wang

Toufeeq

et al. 2020

Arch Insect Biochem Physiol

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Ferritin is a ubiquitous and conserved iron storage protein that plays a significant role in host detoxification, iron storage, and immune response. Although ferritin has been studied in many species, little is known about its role in the Eri‐silkworm (Samia cynthia ricini). In this study, the ferritin light‐chain subunit gene, named ScFerLCH, was identified from S. c. ricini. The full‐length gene, ScFerLCH, was 1,155 bp and encoded a protein consisting of 231 amino acids with a deduced molecular weight of 26.38 kDa. Higher ScFerLCH expression levels were found in the midgut, silk gland, and fat body by quantitative reverse‐transcription polymerase chain reaction and western blot analysis. Injection of Staphylococcus aureus and Pseudomonas aeruginosa could induce upregulation of ScFerLCH in the hemolymph, fat body, and midgut, indicating that ScFerLCH may contribute to the host defense against invading pathogens. In addition, the native ferritin protein was isolated from S. c. ricini by native polyacrylamide gel electrophoresis and its two subunits, ferritin heavy‐chain subunit (ScFerHCH) and ferritin light‐chain subunit (ScFerLCH), were identified by mass spectrometry. Specifically, we found that recombinant ferritin subunits could self‐assemble into a protein complex in vitro; moreover, both recombinant subunits and the protein complex were found to bind different bacteria, including Escherichia coli, P. aeruginosa, S. aureus, and Bacillus subtilis. However, bactericidal tests showed that the protein complex could not inhibit the growth of bacteria directly. Taken together, our results suggest that ScFerritin might play an important role in mediating molecular interaction with pathogens.

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Section: Discussionsupporting

confidence: 85%

Section: Methodsmentioning

confidence: 99%

Identification of FerLCH, isolation of ferritin and functional analysis related to interaction with pathogens in Eri‐silkworm, Samia cynthia ricini

Yang

Wang

Toufeeq

et al. 2020

Arch Insect Biochem Physiol

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“…In higher eukaryotes, the HCH and LCH can be assembled to form heteropolymers. On the contrary, ferritin in the form of a heteropolymer could be depolymerized into the single subunit (Fei et al, ; Theil, ). In recent years, the functions of ferritin subunit have been widely assessed in invertebrates, especially for the functional research of HCH.…”

Section: Discussionmentioning

confidence: 99%

“…In higher vertebrates, ferritin contains two major subunits: heavy (H) and light (L) chain subunits (Harrison & Arosio, ). The insect ferritin is a tetrahedral symmetry consisting of 12H and 12L chain subunits (Fei et al, ). The H chain subunit contains a ferroxidase center that can convert ferrous to ferric iron, whereas the L chain subunit possesses a mineral core to facilitate iron nucleation (Clegg, Fitton, Harrison, & Treffry, ; Santambrogio et al, ).…”

Section: Introductionmentioning

confidence: 99%

Identification and functional analysis of an iron‐binding protein, ferritin heavy chain subunit, from the swallowtail butterfly, Papilio xuthus

Xie

et al. 2019

Arch Insect Biochem Physiol

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Ferritin, which is ubiquitous among all living organisms, plays a crucial role in maintaining iron homeostasis, immune response, and detoxification. In the present research, we identified an iron-binding protein, ferritin heavy chain subunit, from Papilio xuthus and named PxFerHCH. The complete complementary DNA of PxFerHCH was 1,252 bp encoding a sequence of 211 amino acids, which includes an iron-responsive element. Phylogenetic analysis showed that PxFerHCH is clustered with Manduca sexta and Galleria mellonella ferritin heavy chain subunits. Expression levels of PxFerHCH in various tissues were analyzed by reverse transcription quantitative polymerase chain reaction, and the results exhibited that PxFerHCH was expressed in all tissues with the highest expression in the fat body. The relative expression level of PxFerHCH in response to bacterial (Escherichia coli and Staphylococcus aureus) challenges sharply increased by about 12 hr postinfection (hpi) and then decreased at 24 hpi. In addition, the iron-binding capacity and antioxidation activity of recombinant PxFerHCH protein were also investigated. These results reveal that PxFerHCH might play an important role in defense against bacterial infection.

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“…It is also suggested that the expression of BmLHA may be regulated by hormones. Additionally, the BmTCP-1, B. mori heavy and light chain homolog (BmHCH, BmLCH) and B. mori Cytochrome c (BmCytc) genes found to respond to BmNPV infection were regulated by hormones [16,43,44].…”

Section: Discussionmentioning

confidence: 99%

A Novel Digestive Proteinase Lipase Member H-A in Bombyx mori Contributes to Digestive Juice Antiviral Activity against B. mori Nucleopolyhedrovirus

Zhang

Zhu

You

et al. 2020

Insects

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Previous studies have revealed that some proteins in Bombyx mori larvae digestive juice show antiviral activity. Here, based on the label-free proteomics data, BmLipase member H-A (BmLHA) was identified as being involved in the response to BmNPV infection in B. mori larvae digestive juice. In the present study, a gene encoding the BmLHA protein in B. mori was characterized. The protein has an open reading fragment of 999 bp, encoding a predicted 332 amino acid residue-protein with a molecular weight of approximately 35.9 kDa. The phylogenetic analysis revealed that BmLHA shares a close genetic distance with Papilio xuthus Lipase member H-A. BmLHA was highly expressed in the middle part of the B. mori gut, and the expression level increased with instar rising in larvae. There was higher expression of BmLHA in A35 than in P50 strains, and it was upregulated in both A35 and P50 strains, following BmNPV infection. The expression level of VP39 decreased significantly in appropriate recombinant-BmLHA-treated groups compared with the PBS-treated group in B. mori larvae and BmN cells. Meanwhile, overexpression of BmLHA significantly reduced the infectivity of BmNPV in BmN cells. These results indicated that BmLHA did not have digestive function but had anti-BmNPV activity. Taken together, our work provides valuable data for the clarification of the molecular characterization BmLHA and supplements research on proteins of anti-BmNPV activity in B. mori.

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Isolation of ferritin and its interaction with BmNPV in the silkworm, Bombyx mori

Cited by 13 publications

References 45 publications

Identification of FerLCH, isolation of ferritin and functional analysis related to interaction with pathogens in Eri‐silkworm, Samia cynthia ricini

Identification of FerLCH, isolation of ferritin and functional analysis related to interaction with pathogens in Eri‐silkworm, Samia cynthia ricini

Identification and functional analysis of an iron‐binding protein, ferritin heavy chain subunit, from the swallowtail butterfly, Papilio xuthus

A Novel Digestive Proteinase Lipase Member H-A in Bombyx mori Contributes to Digestive Juice Antiviral Activity against B. mori Nucleopolyhedrovirus

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