Identification and Characterization of Molecules Having Anti-microbial Activities from Rabbit Epididymis Using Proteomic Approach

K.V.R, Reddy; Deva, Sukanya; Aranha, Clara; Patgaonkar, ar S.; Bhonde, Gauri

doi:10.4172/1948-5948.s2-002

J Microb Biochem Technol

2011

DOI: 10.4172/1948-5948.s2-002

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Identification and Characterization of Molecules Having Anti-microbial Activities from Rabbit Epididymis Using Proteomic Approach

Reddy K.V.R¹,

Sukanya Deva²,

Clara Aranha³

et al.

Abstract: Antimicrobial peptides (AMPs) are the key components of the host innate immune system and form the first line of host defense against invading pathogens. In the present study attempts were made to identify and characterize AMPs that are synthesized in the epididymal epithelial cells (EPECs) of rabbit, Oryctologus cuniculus. Ex-vivo induction of EPECs with TLR-9 ligand, CpG-ODN-2006 resulted an up-regulation of several AMPs. These molecules were identified in the acid extracts of pooled epididymal tissue homoge… Show more

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Cited by 2 publications

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Effect of Rabbit Epididymal Antimicrobial Peptide, REHbβP, on LPS-Induced Proinflammatory Cytokine Responses in Human Vaginal CellsIn Vitro

Reddy

Sukanya

Patgaonkar

et al. 2012

International Journal of Peptides

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Antimicrobial peptides (AMP's) protect epithelial surfaces including epididymis against pathogens and play a key role in orchestrating various defensive responses. Recently, we have identified one such AMP, rabbit epididymal hemoglobin-β subuit (REHbβP) from the epididymal fluid of rabbit, Oryctologus cuniculus. The demonstration of a protective role of REHbβP in epididymal epithelial cells (EPEC's) led us to investigate: (1) the identification of LPS interactive domain in REHbβP, and (2) whether the REHbβP of rabbit origin mediates vaginal cellular immune responses of another species (human). HeLa-S3, human vaginal epithelial cells (hVECs) were exposed to LPS or the LPS-stimulated cells treated with REHbβP or neutral peptide, nREHbβP. Effect of LPS and cytokines (IL-6 and IL-1α) and chemokines (IL-8, MCP-1) levels was determined in the culture supernatants. In response to the LPS, hVECs synthesized these mediators and the levels were significantly higher than controls. This enhancing effect was ameliorated when the LPS-induced hVECs were treated with REHbβP. Similar results were obtained on NF-κB protein and hBD-1 mRNA expression. Confocal microscopy studies revealed that REHbβP attenuated the LPS-induced internalization of E. coli by macrophages. The chemotaxis studies performed using Boyden chamber Transwell assay, which showed elevated migration of U937 cells when the supernatants of LPS-induced hVECs were used, and the effect was inhibited by REHbβP. REHbβP was found to be localized on the acrosome of rabbit spermatozoa, suggesting its role in sperm protection beside sperm function. In conclusion, REHbβP may have the potential to develop as a therapeutic agent for reproductive tract infections (RTI's).

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Effect of Rabbit Epididymal Antimicrobial Peptide, REHbβP, on LPS-Induced Proinflammatory Cytokine Responses in Human Vaginal CellsIn Vitro

Reddy

Sukanya

Patgaonkar

et al. 2012

International Journal of Peptides

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High-Throughput Identification of Antimicrobial Peptides from Amphibious Mudskippers

You

Bian

et al. 2017

Marine Drugs

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Widespread existence of antimicrobial peptides (AMPs) has been reported in various animals with comprehensive biological activities, which is consistent with the important roles of AMPs as the first line of host defense system. However, no big-data-based analysis on AMPs from any fish species is available. In this study, we identified 507 AMP transcripts on the basis of our previously reported genomes and transcriptomes of two representative amphibious mudskippers, Boleophthalmus pectinirostris (BP) and Periophthalmus magnuspinnatus (PM). The former is predominantly aquatic with less time out of water, while the latter is primarily terrestrial with extended periods of time on land. Within these identified AMPs, 449 sequences are novel; 15 were reported in BP previously; 48 are identically overlapped between BP and PM; 94 were validated by mass spectrometry. Moreover, most AMPs presented differential tissue transcription patterns in the two mudskippers. Interestingly, we discovered two AMPs, hemoglobin β1 and amylin, with high inhibitions on Micrococcus luteus. In conclusion, our high-throughput screening strategy based on genomic and transcriptomic data opens an efficient pathway to discover new antimicrobial peptides for ongoing development of marine drugs.

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Identification and Characterization of Molecules Having Anti-microbial Activities from Rabbit Epididymis Using Proteomic Approach

Cited by 2 publications

References 32 publications

Effect of Rabbit Epididymal Antimicrobial Peptide, REHbβP, on LPS-Induced Proinflammatory Cytokine Responses in Human Vaginal CellsIn Vitro

Effect of Rabbit Epididymal Antimicrobial Peptide, REHbβP, on LPS-Induced Proinflammatory Cytokine Responses in Human Vaginal CellsIn Vitro

High-Throughput Identification of Antimicrobial Peptides from Amphibious Mudskippers

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