Prolyl 4-hydroxlase activity is essential for development and cuticle formation in the human infective parasitic nematode Brugia malayi

Winter, A., McCormack, G., Myllyharju, J. and Page, A. (2013) Prolyl 4-hydroxlase activity is essential for development and cuticle formation in the human infective parasitic nematode Brugia malayi. Journal of Biological Chemistry, 288(3), pp. 1750-1761. (doi: 10.1074/jbc.M112.397604) (PMID:23223450) (PMCID:PMC3548485)

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Publisher's URL: http://dx.doi.org/10.1074/jbc.M112.397604

Abstract

Collagen prolyl 4-hydroxylases (C-P4H) are required for formation of extracellular matrices in higher eukaryotes. These enzymes convert proline residues within the repeat regions of collagen polypeptides to 4-hydroxyproline, a modification essential for the stability of the triple helix. C-P4Hs are most often oligomeric complexes, with enzymatic activity contributed by the α subunits, and the β subunits formed by protein disulfide isomerase (PDI). Here we characterise this enzyme class in the important human parasitic nematode Brugia malayi. All potential C-P4H subunits were identified by detailed bioinformatic analysis of sequence databases, function was investigated both by RNAi in the parasite and heterologous expression in Caenorhabditis elegans, while biochemical activity and complex formation were examined via co-expression in insect cells. Simultaneous RNAi of two B. malayi C-P4H α subunit-like genes resulted in a striking, highly penetrant body morphology phenotype in parasite larvae. This was replicated by single RNAi of a B. malayi C-P4H β subunit-like PDI. Surprisingly however, the B. malayi proteins were not capable of rescuing a C. elegans α subunit mutant, whereas the human enzymes could. In contrast, the B. malayi PDI did functionally complement the lethal phenotype of a C. elegans β subunit mutant. Comparison of recombinant and parasite derived material indicates that enzymatic activity may be dependent on a non-reducible, inter-subunit cross-link, present only in the parasite. We therefore demonstrate that C-P4H activity is essential for development of B. malayi and uncover a novel parasite-specific feature of these collagen biosynthetic enzymes that may be exploited in future parasite control.

Item Type:Articles
Additional Information:his research was originally published in Journal of Biological Chemistry.Winter, A., McCormack, G., Myllyharju, J., and Page, A. (2013) Prolyl 4-hydroxlase activity is essential for development and cuticle formation in the human infective parasitic nematode Brugia malayi. Journal of Biological Chemistry. 2013; 288:1750-1761. © the American Society for Biochemistry and Molecular Biology
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Winter, Dr Alan and McCormack, Ms Gillian and Page, Professor Tony
Authors: Winter, A., McCormack, G., Myllyharju, J., and Page, A.
Subjects:Q Science > QH Natural history > QH345 Biochemistry
College/School:College of Medical Veterinary and Life Sciences > Institute of Infection Immunity and Inflammation
Journal Name:Journal of Biological Chemistry
Journal Abbr.:JBC
Publisher:American Society for Biochemistry and Molecular Biology, Inc.
ISSN:0021-9258
Published Online:06 December 2012
Copyright Holders:Copyright © 2013 The American Society for Biochemistry and Molecular Biology, Inc
First Published:First published in Journal of Biological Chemistry 288:1750-1761
Publisher Policy:Reproduced in accordance with the copyright policy of the publisher
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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
229993The Molecular Enzymology of Collagen Assembly and Post-Translational Modification: a Nematode Model SystemAntony PageMedical Research Council (MRC)G117/476III - BACTERIOLOGY
543261The matrix associated astacin enzymes; novel targets in the control of key GI nematodes of ruminants.Antony PageBiotechnology and Biological Sciences Research Council (BBSRC)BB/I011218/1III - BACTERIOLOGY