Peroxidase catalysed cross-linking of an intrinsically unstructured protein via dityrosine bonds in the oocyst wall of the apicomplexan parasite, Eimeria maxima

Mai, K; Smith, NC; Feng, ZP; Katrib, M; Šlapeta, J; Šlapetova, I; Wallach, MG; Luxford, C; Davies, MJ; Zhang, X; Norton, RS; Belli, SI

Peroxidase catalysed cross-linking of an intrinsically unstructured protein via dityrosine bonds in the oocyst wall of the apicomplexan parasite, Eimeria maxima

Mai, K Smith, NC

Feng, ZP Katrib, M Šlapeta, J Šlapetova, I Wallach, MG Luxford, C Davies, MJ Zhang, X Norton, RS Belli, SI

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Publication Type:: Journal Article
Citation:: International Journal for Parasitology, 2011, 41 (11), pp. 1157 - 1164
Issue Date:: 2011-09-01

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Field	Value	Language
dc.contributor.author	Mai, K	en_US
dc.contributor.author	Smith, NC https://orcid.org/0000-0002-7467-1451	en_US
dc.contributor.author	Feng, ZP	en_US
dc.contributor.author	Katrib, M	en_US
dc.contributor.author	Šlapeta, J	en_US
dc.contributor.author	Šlapetova, I	en_US
dc.contributor.author	Wallach, MG	en_US
dc.contributor.author	Luxford, C	en_US
dc.contributor.author	Davies, MJ	en_US
dc.contributor.author	Zhang, X	en_US
dc.contributor.author	Norton, RS	en_US
dc.contributor.author	Belli, SI https://orcid.org/0000-0003-3845-326X	en_US
dc.date.available	2011-07-02	en_US
dc.date.issued	2011-09-01	en_US
dc.identifier.citation	International Journal for Parasitology, 2011, 41 (11), pp. 1157 - 1164	en_US
dc.identifier.issn	0020-7519	en_US
dc.identifier.uri	http://hdl.handle.net/10453/18170
dc.description.abstract	Apicomplexan parasites such as Eimeria maxima possess a resilient oocyst wall that protects them upon excretion in host faeces and in the outside world, allowing them to survive between hosts. The wall is formed from the contents of specialised organelles - wall-forming bodies - found in macrogametes of the parasites. The presence of dityrosine in the oocyst wall suggests that peroxidase-catalysed dityrosine cross-linking of tyrosine-rich proteins from wall-forming bodies forms a matrix that is a crucial component of oocyst walls. Bioinformatic analyses showed that one of these tyrosine-rich proteins, EmGAM56, is an intrinsically unstructured protein, dominated by random coil (52-70%), with some α-helix (28-43%) but a relatively low percentage of β-sheet (1-11%); this was confirmed by nuclear magnetic resonance and circular dichroism. Furthermore, the structural integrity of EmGAM56 under extreme temperatures and pH indicated its disordered nature. The intrinsic lack of structure in EmGAM56 could facilitate its incorporation into the oocyst wall in two ways: first, intrinsically unstructured proteins are highly susceptible to proteolysis, explaining the several differently-sized oocyst wall proteins derived from EmGAM56; and, second, its flexibility could facilitate cross-linking between these tyrosine-rich derivatives. An in vitro cross-linking assay was developed using a recombinant 42. kDa truncation of EmGAM56. Peroxides, in combination with plant or fungal peroxidases, catalysed the rapid formation of dityrosine cross-linked polymers of the truncated EmGAM56, as determined by western blotting and HPLC, confirming this protein's propensity to form dityrosine bonds. © 2011 Australian Society for Parasitology Inc.	en_US
dc.relation.ispartof	International Journal for Parasitology	en_US
dc.relation.isbasedon	10.1016/j.ijpara.2011.07.001	en_US
dc.subject.classification	Mycology & Parasitology	en_US
dc.subject.mesh	Cell Wall	en_US
dc.subject.mesh	Eimeria	en_US
dc.subject.mesh	Oocysts	en_US
dc.subject.mesh	Peroxidase	en_US
dc.subject.mesh	Tyrosine	en_US
dc.subject.mesh	Protozoan Proteins	en_US
dc.subject.mesh	Cross-Linking Reagents	en_US
dc.subject.mesh	Protein Structure, Tertiary	en_US
dc.subject.mesh	Biocatalysis	en_US
dc.title	Peroxidase catalysed cross-linking of an intrinsically unstructured protein via dityrosine bonds in the oocyst wall of the apicomplexan parasite, Eimeria maxima	en_US
dc.type	Journal Article
utslib.citation.volume	11	en_US
utslib.citation.volume	41	en_US
utslib.for	0605 Microbiology	en_US
utslib.for	0608 Zoology	en_US
utslib.for	0707 Veterinary Sciences	en_US
pubs.embargo.period	Not known	en_US
pubs.organisational-group	/University of Technology Sydney
pubs.organisational-group	/University of Technology Sydney/Faculty of Science
pubs.organisational-group	/University of Technology Sydney/Faculty of Science/School of Life Sciences
utslib.copyright.status	closed_access
pubs.issue	11	en_US
pubs.publication-status	Published	en_US
pubs.volume	41	en_US

Abstract:

Apicomplexan parasites such as Eimeria maxima possess a resilient oocyst wall that protects them upon excretion in host faeces and in the outside world, allowing them to survive between hosts. The wall is formed from the contents of specialised organelles - wall-forming bodies - found in macrogametes of the parasites. The presence of dityrosine in the oocyst wall suggests that peroxidase-catalysed dityrosine cross-linking of tyrosine-rich proteins from wall-forming bodies forms a matrix that is a crucial component of oocyst walls. Bioinformatic analyses showed that one of these tyrosine-rich proteins, EmGAM56, is an intrinsically unstructured protein, dominated by random coil (52-70%), with some α-helix (28-43%) but a relatively low percentage of β-sheet (1-11%); this was confirmed by nuclear magnetic resonance and circular dichroism. Furthermore, the structural integrity of EmGAM56 under extreme temperatures and pH indicated its disordered nature. The intrinsic lack of structure in EmGAM56 could facilitate its incorporation into the oocyst wall in two ways: first, intrinsically unstructured proteins are highly susceptible to proteolysis, explaining the several differently-sized oocyst wall proteins derived from EmGAM56; and, second, its flexibility could facilitate cross-linking between these tyrosine-rich derivatives. An in vitro cross-linking assay was developed using a recombinant 42. kDa truncation of EmGAM56. Peroxides, in combination with plant or fungal peroxidases, catalysed the rapid formation of dityrosine cross-linked polymers of the truncated EmGAM56, as determined by western blotting and HPLC, confirming this protein's propensity to form dityrosine bonds. © 2011 Australian Society for Parasitology Inc.

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http://hdl.handle.net/10453/18170