바지락(Ruditapes philippinarum) 아가미로부터항균 펩타이드의 탐색 및 정제
Screening and Purification of an Antimicrobial Peptide from the Gill of the Manila clam Ruditapes philippinarum
2
조회
0
다운로드
7
인용
0
참고문헌
키워드
초록
This study screened the biological activity of an acidified gill extract of the Manila clam Ruditapes philippinarum including antimicrobial, hemolytic, membrane permeabilization, and DNA-binding activity, and purified the antimicrobial material. The acidified gill extract showed potent antimicrobial activity against Bacillus subtilis and Escherichia coli without significant hemolytic activity, but showed no membrane permeabilization or DNA-binding ability. An antimicrobial material was purified from the acidified gill extract using C18 reversed-phase and cation-exchange highperformance liquid chromatography (HPLC). Treatment of the purified material with trypsin completely abolished all of the antibacterial activity against Bacillus subtilis, suggesting that the purified material is a proteinaceous antibiotic. The molecular weight of the purified material was 2571.9 Da, but no primary structural information was obtained due to N-terminal blocking. A future study should confirm the primary structure. Our results suggest that the Manila clam gill contains proteinaceous antibiotics that have a role in first-line defense. This information could be used to better understand the Manila clam innate immune system.
Abstract
This study screened the biological activity of an acidified gill extract of the Manila clam Ruditapes philippinarum including antimicrobial, hemolytic, membrane permeabilization, and DNA-binding activity, and purified the antimicrobial material. The acidified gill extract showed potent antimicrobial activity against Bacillus subtilis and Escherichia coli without significant hemolytic activity, but showed no membrane permeabilization or DNA-binding ability. An antimicrobial material was purified from the acidified gill extract using C18 reversed-phase and cation-exchange highperformance liquid chromatography (HPLC). Treatment of the purified material with trypsin completely abolished all of the antibacterial activity against Bacillus subtilis, suggesting that the purified material is a proteinaceous antibiotic. The molecular weight of the purified material was 2571.9 Da, but no primary structural information was obtained due to N-terminal blocking. A future study should confirm the primary structure. Our results suggest that the Manila clam gill contains proteinaceous antibiotics that have a role in first-line defense. This information could be used to better understand the Manila clam innate immune system.
참고문헌 (35)
- Alfred RL, "The antimicrobial domains of wheat puroindolines are cell-penetrating peptides with possible intracellular mechanism of action", PLoS One, 8, pp.e75488~ (2013)
- Andreu D, "Animal antimicrobial peptides: an overview", Biopolymers, 147, pp.415~433 (1998)
- Charlet M, "Innate immunity: Isolation of several cysteine- rich antimicrobial peptides from the blood of a mollusk, Mytilus edulis", J Biol Chem, 271, pp.21808~21813 (1996)
- Destoumieux D, "Penaeidins, a new family of antimicrobial peptides isolated from the shrimp Penaeus vannamei (Decapoda)", J Biol Chem, 272, pp.28398~28406 (1997)
- De Zoysa M, "Abhisin: a potential antimicrobial peptide derived from histone H2A of disk abalone (Haliotis discus discus)", Fish Shellfish Immunol, 27, pp.639~646 (2009)
- Gueguen Y, "Characterization of a defensin from the oyster Crassostrea gigas. Recombinant production, folding, solution structure, antimicrobial activities, and gene expression", J Biol Chem, 281, pp.313~323 (2006)
- Robert E.W Hancock, "The role of cationic antimicrobial peptides in innate host defences", Trends in Microbiology, 8(9), pp.402~410 (2000), Elsevier BV[DOI]
- Hsu CH, "Structural and DNA-binding studies on the bovine antimicrobial peptide, indolicidin, evidence for multiple conformations involved in binding to membranes and DNA", Nucl Acids Res, 33, pp.4053~4064 (2005)
- Iijima R, "A novel antimicrobial peptide from the sea hare Dolabella auricularia", Dev Comp Immunol, 27, pp.305~311 (2003)
- Izadpanah A, "Antimicrobial peptides", J Am Acad Dermatol, 52, pp.381~390 (2005)
- Kang YS, "Analysis of EST and lectin expression in hemocytes of Manila clams (Ruditapes phylippinarum) (Bivalvia: Mollusca) infected with Perkinsus olseni", Dev Comp Immunol, 30, pp.1119~1131 (2006)
- Khoo L1, "Callinectin, an antibacterial peptide from blue crab, Callinectes sapidus, hemocytes", Mar Biotechnol (NY), 1, pp.44~51 (1999)
- Lehrer RI, "Ultrasensitive assay for endogenous antimicrobial polypeptides", J Immunol Methods, 137, pp.167~173 (1991)
- Li Y, "The role of antimicrobial peptides in cardiovascular physiology and disease", Biochem Biophys Res Commun, 390, pp.363~367 (2009)
- Mitta G, "Mytilin B and MGD2, two antimicrobial peptides of marine mussels: gene structure and expression analysis", Dev Comp Immunol, 24, pp.381~393 (2000)
- Mitta G, "Myticin, a novel cysteine-rich antimicrobial peptide isolated from haemocytes and plasma of the mussel Mytilus galloprovincialis", Eur J Biochem, 265, pp.71~78 (1999)
- Miyata T, "Antimicrobial peptides, isolated from horseshoe crab hemocytes, tachyplesin II, and polyphemusins I and II: chemical structures and biological activity", J Biochem (Tokyo), 106, pp.663~668 (1989)
- Nam BH, "Functional analysis of Pacific oyster (Crassostrea gigas) β-thymosin: Focus on antimicrobial activity", Fish Shellfish Immunol, 45, pp.167~174 (2015)
- Ovchinnikova TV, "Purification and primary structure of two isoforms of arenicin, a novel antimicrobial peptide from marine polychaeta Arenicola marina", FEBS Lett, 577, pp.209~214 (2004)
- Park KI, "Report on the occurrence of brown ring disease (BRD) in Manila clam, Ruditapes philippinarum, on the west coast of Korea", Aquaculture, 255, pp.610~613 (2006)
- Relf JM, "Purification and characterization of a cystein-rich 11.5-kDa antibacterial protein from the granular haemocytes of the shore crab, Carcinus maenas", Eur J Biochem, 264, pp.350~357 (1999)
- Saito T, "A novel big defensin identified in horseshoe crab hemocytes: isolation, amino acid sequence, and antibacterial activity", J Biochem (Tokyo), 117, pp.1131~1137 (1995)
- Seo JK, "Purification of a novel arthropod defensin from the American oyster, Crassostrea virginica", Biochem Biophys Res Commun, 33, pp.1998~2004 (2005)
- Seo JK, "Purification and antimicrobial function of ubiquitin isolated from the gill of Pacific oyster, Crassostrea gigas", Mol Immunol, 53, pp.88~98 (2013)
- Seo JK, "Antimicrobial function of the GAPDH-related antimicrobial peptide in the skin of skipjack tuna, Katsuwonus pelamis", Fish Shellfish Immunol, 36, pp.571~581 (2014)
- Seo JK, "cgMolluscidin, a novel dibasic residue repeat rich antimicrobial peptide, purified from the gill of the Pacific oyster, Crassostrea gigas", Fish Shellfish Immunol, 35, pp.480~488 (2013)
- Shai Y, "qMode of action of membrane active antimicrobial peptides", Biopolymers, 66, pp.236~248 (2002)
- Silphaduang U, "Peptide antibiotics in mast cells of fish", Nature, 414, pp.268~269 (2001)
- Skerlavaj B, "Rapid membrane permeabilization and inhibition of vital functions of Gramnegative bacteria by bactenecins", Infect Immun, 58, pp.3724~3730 (1990)
- M. Stark, "Cationic Hydrophobic Peptides with Antimicrobial Activity", Antimicrobial Agents and Chemotherapy, 46(11), pp.3585~3590 (2002), American Society for Microbiology[DOI]
- Tincu JA, "Antimicrobial peptides from marine invertebrates", Antimicrob Agents Chemother, 48, pp.3645~3654 (2004)
- Wim van t Hof, "Antimicrobial Peptides: Properties and Applicability", Biological Chemistry, 382(4), pp.597~619 (2001), Walter de Gruyter GmbH[DOI]
- Zasloff M, "Antimicrobial peptides of multicellular organisms", Nature, 415, pp.389~395 (2002)
- Zhang L, "A defensin from clam Venerupis philippinarum: Molecular characterization, localization, antibacterial activity, and mechanism of action", Dev Comp Immunol, 51, pp.29~38 (2015)
- Zhao J, "Molecular characterization of a novel big defensin from clam, Venerupis philippinarum", PLoS One, 5, pp.e13480~ (2010)
