Identification and characterization of an 18.4kDa antimicrobial truncation from shrimp Litopenaeus vannamei hemocyanin upon Vibrio parahaemolyticus infection

Yuan, Wen; Zhan, Shixiong; Huang, He; Zhong, Min; Chen, Jiehui; You, Cuihong; Wang, Fan; Zhang, Yueling

doi:10.1016/j.fsi.2016.08.002

Cited by 40 publications

(21 citation statements)

References 29 publications

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“…A 19-mer peptide, HP (2-20), derived from the N-terminal region of Helicobacter pylori ribosomal protein L1 (RPL1), was shown to possess broad-spectrum antimicrobial activities (Park and Hahm 2005). Similarly, hemocyanin, the most abundant protein in arthropods, is reported to also generate various AMPs (Lee et al 2003; Destoumieux-Garzon et al 2001; Wen et al 2016). …”

Section: Introductionmentioning

confidence: 99%

Prediction and characterization of a novel hemocyanin-derived antimicrobial peptide from shrimp Litopenaeus vannamei

et al. 2018

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Hemocyanin, the multifunctional glycoprotein in the hemolymph of invertebrates, can generate various antimicrobial peptides (AMPs). Given the rising interest in the use of natural therapeutic agents such as AMPs, alternative and more efficient methods for their generation are being explored. In this work, free online software was first applied to predict the generation of antimicrobial peptides from the large subunit of Litopenaeus vannamei hemocyanin. Twenty potential antimicrobial peptides ranging from 1.5 to 1.9 kDa were predicted, five of which had α-helical structures and were selected for antibacterial activity testing. The results indicated that these five peptides had antibacterial activity against seven different bacteria. Of the five peptides, one peptide, designated L1, had the strongest antibacterial activity against both Gram-negative and Gram-positive bacteria. Moreover, CD and NMR data showed that L1 had both α-helical and β-turns structural composition, and that these structures were essential for L1’s antibacterial activity. Furthermore, SEM analysis revealed that peptide L1 had broad-spectrum activity against both Gram-positive and Gram-negative bacteria, as it could destroy the bacterial cell walls and kill the bacteria. Thus, L1 is a very potent antimicrobial peptide that can be exploited and used in antibacterial therapeutics.Electronic supplementary materialThe online version of this article (10.1007/s00726-018-2575-x) contains supplementary material, which is available to authorized users.

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Section: Introductionmentioning

confidence: 99%

Prediction and characterization of a novel hemocyanin-derived antimicrobial peptide from shrimp Litopenaeus vannamei

et al. 2018

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“…We recently found that shrimp seem to have different immune defensive strategies against different virulent pathogens, with the N and C termini of small subunit hemocyanin having different functions, such as agglutinative activity toward E. coli K12 and chicken erythrocytes . Of particular note is the fact that, a 18.4 kDa peptide in the C terminus of small subunit hemocyanin, which we previously found to have strong agglutination and antibacterial activities against pathogenic bacteria, was found in this study to contain three of the glycosylation sites (Thr‐537, Ser‐539, and Thr‐542) . Even though arthropods can produce mucin‐type O‐glycans, there was a Hex (for glucose, mannose, galactose) type glycan on site Thr‐537, which might be a new type glycan in arthropods.…”

Section: Discussionmentioning

confidence: 48%

“…Aside from this, we also previously showed that an 18.4 kDa peptide on the C terminus of small subunit hemocyanin (double underlined in Fig. B) had strong agglutination and antibacterial activities against pathogenic bacteria . Based on these, we selected three potential immune‐related glycosylation sites on this C‐terminal peptide to generate a mutant of small subunit hemocyanin, mut‐rHMCs, by replacing threonine (Thr) and serine (Ser) residues at positions 537, 542, and 539 with alanine (Ala) (Fig.…”

Section: Resultsmentioning

confidence: 89%

Identification and characterization of glycosylation sites on Litopenaeus vannamei hemocyanin

Zhang

Aweya

et al. 2019

FEBS Letters

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The respiratory glycoprotein hemocyanin has been implicated in immune‐related functions. Using lectin blotting, we show that the binding of shrimp (Litopenaeus vannamei) hemocyanin to concanavalin A decreases markedly with O‐glycosidase treatment but not with PNGase F. Twelve O‐glycosylation sites, three on the large hemocyanin subunit and nine on the small hemocyanin subunit (HMCs), were identified by LC‐MS/MS. Importantly, when the glycosylation sites at Thr‐537, Ser‐539, and Thr‐542 on the C terminus of HMCs were replaced with alanine, the resultant mutant hemocyanin had reduced carbohydrate content, coupled with a fourfold reduction in bacterial agglutination and 0.2‐fold reduction in antibacterial activities toward Vibrio parahaemolyticus and Staphylococcus aureus. These results suggest that the glycosylation sites on shrimp hemocyanin are closely related to its immunological functions.

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“…Hemocyanin has exhibited excellent antimicrobial (15)(16)(17), anti-tumor (18) and immune enhancement effects (19). In the present study, hemocyanin was used as an additive in T cell culture in vitro.…”

Section: Discussionmentioning

confidence: 92%

The effects of hemocyanin on T cells cultured inï¿½vitro

Liu

Chen

et al. 2017

Oncol Lett

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Abstract. As a broad-spectrum antibiotic, gentamicin is used extensively in T cell culturing in vitro, but preliminary studies have identified that T cell activity is significantly affected by gentamicin. In the present study, the hemocyanin from Litopenaeus vannamei (L. vannamei) was selected as an additive for T cell cultures in vitro. Compared with those in the control group, the cell quantity exhibited no significant difference, and the formation rate of cell colony increased gradually with increases in the hemocyanin concentration. Additionally, flow cytometry assays identified that cluster of differentiation (CD)3 + CD4 + and CD4 + CD25 + T cells in the 0.2 µg/ml hemocyanin (Hem 3) group were all significantly increased. Furthermore, cell cycle analysis demonstrated that T cells in the G0/G1 phase were significantly decreased in the Hem 3 group compared with in the control, 0.05 µg/ml (Hem 1) and 0.1 µg/ml (Hem 2) groups, and cells in the S phase were significantly elevated in the Hem 3 group compared with in the control and Hem 1 groups. In addition, MTT analysis indicated that the cytotoxicity of T cells towards HepG2 cells was significantly increased in the Hem 3 group compared with in the control, Hem 1 and Hem 2 groups. Taken together, the present study identified that hemocyanin may improve the proliferation and cytotoxicity of T cells, and the results supported the use of hemocyanin in T cell adoptive immunotherapy.

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Identification and characterization of an 18.4kDa antimicrobial truncation from shrimp Litopenaeus vannamei hemocyanin upon Vibrio parahaemolyticus infection

Cited by 40 publications

References 29 publications

Prediction and characterization of a novel hemocyanin-derived antimicrobial peptide from shrimp Litopenaeus vannamei

Prediction and characterization of a novel hemocyanin-derived antimicrobial peptide from shrimp Litopenaeus vannamei

Identification and characterization of glycosylation sites on Litopenaeus vannamei hemocyanin

The effects of hemocyanin on T cells cultured inï¿½vitro

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