RRC ID 46468
著者 Winter AD, McCormack G, Myllyharju J, Page AP.
タイトル Prolyl 4-hydroxlase activity is essential for development and cuticle formation in the human infective parasitic nematode Brugia malayi.
ジャーナル J Biol Chem
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 final triple helix. C-P4H 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 characterize 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, whereas 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 covalent 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.
巻・号 288(3)
ページ 1750-61
公開日 2013-1-18
DOI 10.1074/jbc.M112.397604
PII S0021-9258(20)46589-5
PMID 23223450
PMC PMC3548485
MeSH Animals Brugia malayi / enzymology* Brugia malayi / genetics Caenorhabditis elegans / enzymology Caenorhabditis elegans / genetics Collagen / metabolism* Computational Biology Gene Expression Gene Expression Regulation Genetic Complementation Test Helminth Proteins / chemistry Helminth Proteins / genetics* Helminth Proteins / metabolism Humans Mutation Procollagen-Proline Dioxygenase / chemistry Procollagen-Proline Dioxygenase / genetics* Procollagen-Proline Dioxygenase / metabolism Protein Subunits / chemistry Protein Subunits / genetics* Protein Subunits / metabolism RNA, Small Interfering / genetics
IF 4.238
引用数 12
WOS 分野 BIOCHEMISTRY & MOLECULAR BIOLOGY
リソース情報
線虫 tm0689