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)
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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: | Murray, Ms Linda and Britton, Professor Collette and Geldhof, Prof Peter |
| 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 > School of Infection & Immunity |
| Journal Name: | Trends in Parasitology |
| Publisher: | Elsevier (Cell Press) |
| ISSN: | 1471-4922 |
| ISSN (Online): | 1471-5007 |
| Published Online: | 21 March 2007 |
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