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Bioscience Methods
BM 2011, Vo.2, No.4, 21-30
http://bm.sophiapublisher.com
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Research Article Open Access
Isolation, Biochemical Characterization and Anti-adhesion Property of Mucin from
the Blue Blubber Jellyfish (
Catostylus mosaicus
)
Roger Pearson
1
, Ross Tellam
1
, B. Xu
2
, Z. Zhao
2,3
, Mark Willcox
2,3
Kritaya Kongsuwan
1
1. CSIRO Livestock Industries, Level 6 Queensland Bioscience Precinct, 306 Carmody Rd., St Lucia QLD 4067, Australia
2. Brien Holden Vision Institute, Level 4 North Wing, Rupert Myers Building, Gate 14 Barker Street, University of New South Wales, Kensington, NSW 2052,
Australia
3. The School of Optometry and Vision Science, University of New South Wales, Kensington, NSW 2052, Australia
Corresponding author email: Kritaya.Kongsuwan@csiro.au
Author
Bioscience Methods 2011, Vol.2 No.4 DOI:10.5376/bm.2011.02.0004
Received: 18 May, 2011
Accepted: 23 May, 2011
Published: 03 Jun., 2011
This is an open access article published under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and
reproduction in any medium, provided the original work is properly cited.
Preferred citation for this article as:
Pearson et al., 2011, Isolation, biochemical characterization and anti-adhesion property of mucin from the blue blubber jellyfish (
Catostylus mosaicus
),
Bioscience Methods, doi:10.5376/bm.2011.02.0004
Abstract
Mucins are large glycoproteins that have been identified as the main component of various biological gels and
lubricants. Their compositions contribute to the viscoelastic properties of mucus secretions. Gel-forming mucins have now been
identified in a variety of organisms from marine molluscs to humans and are thought to cover and protect epithelial cells from
attachment and entry of pathogens. We employed a new approach using a combination of tryptic digestion and the fractionation by
hydrophobic interaction chromatography to enable the isolation and purification of mucins from the Blue Blubber jellyfish,
Catostylus mosaicus.
The purified proteins were stained with Alcian blue indicating extensive glycosylation. Amino acid composition
analysis of the purified protein found that it was enriched for Ala, Glu, Thr, Pro and Val residues (together constituting 93 mole % of
the protein), which is typical for mucins. Consistent with this, monosaccharide composition analysis revealed extensive O-linked
oligosaccharides with N-acetylgalactosamine (GalNAc), galactose (Gal) and N-acetylglucosamine (GlcNAc) as major
monosaccharide constituents. The purified
C. mosaicus
mucins inhibited the attachment of an ocular
Pseudomonas aeruginosa
(
Pa
)
isolate (Paer6294) to human corneal epithelial (HCE) cells grown
in vitro
.
Keywords
Jellyfish; Mucin; Anti-adherence; Oligosaccharides;
Pseudomonas aeruginosa
Background
Jellyfish are found throughout the oceans and seas of
the world, ranging from the waters of the Arctic and
Antarctic through to the equator. At certain times of
the year they swarm in their millions causing
problems for the fishing, power generation, shipping
and tourism industries. In Eastern and Northern
seaboards of Australia the most common jellyfish is
the edible Rhizostome,
Catostylus mosaicus
(commonly
referred to as the Blue Blubber jellyfish).
Mucins belong to a family of glycoproteins that
contains a large amount of O-linked oligosaccharides.
These oligosaccharides are responsible for the
viscoelastic properties of mucous secretions and for
providing protection of the exposed epithelial surfaces
from dehydration, microbial invasion and physical
injuries (Vanklinken et al., 1995; Watanabe 2002).
Secreted mucins present in the ocular tear-film of
humans have been reported to help maintain fluid
viscosity and facilitate lubrication of the eye by
retarding fluid evaporation and anchoring the aqueous
tear-film to the underlying cornea and conjunctival
surfaces (Argüeso and Gipson 2001; Davidson and
Kuonen 2004; Gipson et al., 2004). In addition,
mucins in tear fluid also protect against pathogens by
acting as decoy receptors for the binding of pathogens
to corneal epithelial surfaces (Mantelli and Argueso
2008). Human tear fluid which contains a mixture of
secreted and shed membrane-associated mucins, was
found to be protective against
Pseudomonas
aeruginosa
(
Pa
)’s colonisation in both cultured
corneal epithelial cells (Fleiszig et al., 2003) and in an
animal model where corneas were challenged with
cytotoxic
Pa
(Kwong et al., 2007). Epidemiologic
evidence suggested that conjunctiva mucin secretion