Expression and purification of an active cysteine protease of Haemonchus contortus using Caenorhabditis elegans

Murray, L., Geldhof, P., Clark, D., Knox, D. P. and Britton, C. (2007) Expression and purification of an active cysteine protease of Haemonchus contortus using Caenorhabditis elegans. Trends in Parasitology, 37(10), pp. 1117-1125. (doi: 10.1016/j.ijpara.2007.02.012) (PMID:17451718)

Full text not currently available from Enlighten.

Abstract

Many proteolytic enzymes of parasitic nematodes have been identified as possible targets of control. Testing these as vaccine or drug targets is often difficult due to the problems of expressing proteases in a correctly folded, active form in standard expression systems. In an effort to overcome these difficulties we have tested Caenorhabditis elegans as an expression system for a Haemonchus contortus cathepsin L cysteine protease, Hc-CPL-1. Recombinant Hc-CPL-1 with a polyhistidine tag added to the C-terminal was expressed in an active and glycosylated form in C. elegans. Optimal expression was obtained expressing Hc-cpl-1 under control of the promoter of the homologous C. elegans cpl-1 gene. The recombinant protein was purified from liquid cultures by nickel chelation chromatography in sufficient amounts for vaccination studies to be carried out. This study provides proof of principle that active, post-translationally modified parasitic nematode proteases can be expressed in C. elegans and this approach can be extended for expression of known protective antigens.

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Geldhof, Prof Peter and Britton, Dr Collette and Murray, Ms Linda
Authors: Murray, L., Geldhof, P., Clark, D., Knox, D. P., and Britton, C.
College/School:College of Medical Veterinary and Life Sciences
College of Medical Veterinary and Life Sciences > Institute of Infection Immunity and Inflammation
Journal Name:Trends in Parasitology
Publisher:Elsevier (Cell Press)
ISSN:1471-4922
ISSN (Online):1471-5007
Published Online:21 March 2007

University Staff: Request a correction | Enlighten Editors: Update this record

Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
351421Adaptation of caenorhabditis elegans technology to identification and expression of parasite target genesCollette BrittonBiotechnology and Biological Sciences Research Council (BBSRC)S20387Infection Immunity and Inflammation Life Sciences