Biopolymers from marine invertebrates. X. Mode of action of an antibacterial glycoprotein, aplysianin E, from eggs of a sea hare, Aplysia kurodai.

Kisugi, Jun; Ohye, Hirokazu; Kamiya, Hisao; Yamazaki, Masatoshi

doi:10.1248/cpb.37.3050

Cited by 25 publications

(12 citation statements)

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“…Unlike previous reports of similar proteins isolated from sea hares (Kisugi et al, 1989;Yamazaki et al, 1990;Melo et al, 1998Melo et al, , 2000, we demonstrate that substrates determined the bactericidal or bacteriostatic effects of escapin. Bacteria cultured in M9 and glucose medium without high levels (50·mmol·l -1 ) of selected amino acids or without Tryptone Peptone supplements were not killed (Figs·7, 8).…”

Section: Discussioncontrasting

confidence: 99%

“…The existence of persister cells in our cultures of E. coli may explain why several sea hare homologues of escapin, including aplysianin-E (Kisugi et al, 1989), aplysianin-P (Yamazaki et al, 1990) and dactylomelin-P isolated from purple fluid of Aplysia dactylomela (Melo et al, 2000), have been characterized as bacteriostatic and not bactericidal, whereas we found escapin to be bactericidal. To test this idea, we extracted and purified dactylomelin-P from Aplysia dactylomela and found that it was also bactericidal under our assay conditions; K.-C. Ko and H. Yong, unpublished data).…”

Section: Discussionmentioning

confidence: 68%

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Cloning, characterization and expression of escapin, a broadly antimicrobial FAD-containing l-amino acid oxidase from ink of the sea hare Aplysia californica

Yang

Johnson

et al. 2005

Journal of Experimental Biology

108

122

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SUMMARY A 60 kDa monomeric protein isolated from the defensive purple ink secretion of the sea hare Aplysia californica was cloned and sequenced, and is the first sea hare antimicrobial protein to be functionally expressed in E. coli. Sequence analysis suggested that this protein is a flavin-containing l-amino acid oxidase (LAAO), with one predicted potential glycosylation site, although the glycosylation could not be experimentally confirmed. This protein, which we call `escapin', has high sequence similarity to several other gastropod proteins. Escapin was verified by NMR, mass spectroscopy and HPLC to have FAD as its flavin cofactor. Escapin's antimicrobial effects, bacteriostasis and bactericidal, were determined using a combination of two assays: (1) incubation of bacteria on solid media followed by assessment of inhibition by direct observation of zones of inhibition or by turbidity measurements; and (2) incubation of bacteria in liquid media followed by counting viable colonies after growing on agar plates. Native escapin inhibited the growth of Gram-positive and Gram-negative bacteria, including marine bacteria (Vibrio harveyiiand Staphylococcus aureus) and pathogenic bacteria(Staphylococcus aureus, Streptococcus pyogenes and Pseudomonas aeruginosa). Escapin also inhibited the growth of yeast and fungi, with different efficacies. Escapin's antimicrobial activity was concentration dependent and did not decrease when stored for more than 5 months at room temperature. Escapin was bacteriostatic and not bactericidal in minimal media (e.g. salt media) with glucose, yeast extract, and a mixture of 20 amino acids each at 50 μmol l-1, but was bactericidal in media enriched with Tryptone Peptone. Escapin was also strongly bactericidal in media with l-lysine at concentrations as low as 3 mmol l-1 and slightly bactericidal in 50 mmol l-1l-arginine, but not in most other amino acids even at 50 mmol l-1. Escapin had high oxidase activity (producing hydrogen peroxide) with either l-arginine or l-lysine as a substrate and little to no oxidase activity with other l-amino acids. Hydrogen peroxide alone (without escapin or amino acids) was strongly bacteriostatic but poorly bactericidal, similar in this respect to l-arginine but different from l-lysine in the presence of escapin. Together these results suggest that there are multiple mechanisms to escapin's antimicrobial effects, with bacteriostasis resulting largely or entirely from the effects of hydrogen peroxide produced by escapin's LAAO activity, but bactericidal effects resulting from lysine-dependent mechanisms not directly involving hydrogen peroxide. Recombinant escapin expressed in bacteria was also active against Gram-positive and Gram-negative bacteria,suggesting that glycosylation is not essential for antimicrobial activity.

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Section: Discussioncontrasting

confidence: 99%

Section: Discussionmentioning

confidence: 68%

Cloning, characterization and expression of escapin, a broadly antimicrobial FAD-containing l-amino acid oxidase from ink of the sea hare Aplysia californica

Yang

Johnson

et al. 2005

Journal of Experimental Biology

108

122

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“…The mode of action of an antibacterial glycoprotein was designated and studied by Kisugi et al [16] ,i.e. aplysianin E against a range of bacteria.…”

Section: Aplysianin Ementioning

confidence: 99%

RETRACTED: Antimicrobial secondary metabolites from marine gastropod egg capsules and egg masses

Kaviarasan

Siva

Yogamoorthi

2012

Asian Pacific Journal of Tropical Biomedicine

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“…In addition, there was no apparent decrease in turbidity (data not shown), suggesting that aplysianin-A has no bacteriolytic activity. These facts indicate that aplysianin-A was ~c~ ~~~~laxgm~llg~~ so not bacteriocidal, but bacteriostatic like aplysianin-E, an antibacterial glycoprotein in the egg mass of A. kurodai [1,17].…”

Section: <___mentioning

confidence: 95%

Molecular cloning of the defense factor in the albumen gland of the sea hare Aplysia kurodai

1995

FEBS Letters

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Aplysianin-A, an antibacterial glycoprotein in the albumen gland of the sea hare Aplysia kurodai, inhibited the growth of both Gram-positive and Gram-negative bacteria. Aplysianin-A cDNA clones were isolated from an albumen gland cDNA library. Sequence analysis reveals that aplysianin-A is produced as a precursor protein of 556 amino acid residues with a signal peptide of 19 amino acid residues and contains 6 potential N-glycosylation sites. Aplysianin-A mRNA was expressed tissue-specifically in the albumen gland. Homology search reveals that aplysianin-A has a 50% overall amino acid sequence homology to achacin, an antibacterial glycoprotein of the giant African snail Achatina fulica.

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Biopolymers from marine invertebrates. X. Mode of action of an antibacterial glycoprotein, aplysianin E, from eggs of a sea hare, Aplysia kurodai.

Cited by 25 publications

References 0 publications

Cloning, characterization and expression of escapin, a broadly antimicrobial FAD-containing l-amino acid oxidase from ink of the sea hare Aplysia californica

Cloning, characterization and expression of escapin, a broadly antimicrobial FAD-containing l-amino acid oxidase from ink of the sea hare Aplysia californica

RETRACTED: Antimicrobial secondary metabolites from marine gastropod egg capsules and egg masses

Molecular cloning of the defense factor in the albumen gland of the sea hare Aplysia kurodai

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