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Cloning and characterization of an orphan seven transmembrane receptor from Schistosoma mansoni

Published online by Cambridge University Press:  23 August 2007

M. S. PEARSON ,
D. P. McMANUS ,
D. J. SMYTH ,
M. K. JONES ,
A. M. SYKES  and
A. LOUKAS
M. S. PEARSON*
Affiliation:
Division of Infectious Diseases and Immunology, Queensland Institute of Medical Research and Australian Centre for International and Tropical Health and Nutrition, The University of Queensland, Brisbane, Queensland, Australia
D. P. McMANUS
Affiliation:
Division of Infectious Diseases and Immunology, Queensland Institute of Medical Research and Australian Centre for International and Tropical Health and Nutrition, The University of Queensland, Brisbane, Queensland, Australia
D. J. SMYTH
Affiliation:
Division of Infectious Diseases and Immunology, Queensland Institute of Medical Research and Australian Centre for International and Tropical Health and Nutrition, The University of Queensland, Brisbane, Queensland, Australia
M. K. JONES
Affiliation:
Division of Infectious Diseases and Immunology, Queensland Institute of Medical Research and Australian Centre for International and Tropical Health and Nutrition, The University of Queensland, Brisbane, Queensland, Australia
A. M. SYKES
Affiliation:
Division of Infectious Diseases and Immunology, Queensland Institute of Medical Research and Australian Centre for International and Tropical Health and Nutrition, The University of Queensland, Brisbane, Queensland, Australia
A. LOUKAS*
Affiliation:
Division of Infectious Diseases and Immunology, Queensland Institute of Medical Research and Australian Centre for International and Tropical Health and Nutrition, The University of Queensland, Brisbane, Queensland, Australia
*
*Corresponding authors: Tel: 61 7 3845 3702. Fax: 61 7 3845 3507. E-mail: alex.loukas@qimr.edu.au or mark.pearson@qimr.edu.au
*Corresponding authors: Tel: 61 7 3845 3702. Fax: 61 7 3845 3507. E-mail: alex.loukas@qimr.edu.au or mark.pearson@qimr.edu.au
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Summary

A partial cDNA sequence was obtained from the human blood fluke, Schistosoma mansoni using a signal sequence trap approach. The full-length cDNA was cloned and termed Sm-7TM. The corresponding open reading frame had 7 membrane spanning domains and shared identity with a small, novel group of seven transmembrane (7TM) receptors from vertebrates and invertebrates, including the human ee3 receptor – a heptahelical protein implicated in neuronal signalling. Phylogenetic analysis of this novel family showed that the Sm-7TM ORF formed a clade with exclusively invertebrate sequences. Based on topology modelling with ee3, Sm-7TM was predicted to possess an intracellular C-terminal tail, which was expressed as a soluble thioredoxin fusion protein (Sm-7TMC) in Escherichia coli and purified using metal ion-affinity chromatography. A polyclonal antiserum against this domain was used to detect Sm-7TM in detergent-soluble parasite extracts and to immunolocalize the receptor to the tegument of adult S. mansoni.

Keywords

Schistosoma mansoniseven transmembrane receptortegument
Type
Research Article
Information
Parasitology , Volume 134 , Issue 14 , December 2007 , pp. 2001 - 2008
Copyright
Copyright © Cambridge University Press 2007

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