Journal of Biochemistry Advance Access originally published online on December 14, 2006
Journal of Biochemistry 2007 141(1):47-55; doi:10.1093/jb/mvm009
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© 2006 The Japanese Biochemical Society.
Ligand-binding Activity and Expression Profile of Annexins in Caenorhabditis elegans
1Graduate School of Humanities and Sciences, Ochanomizu University, 2-1-1 Otsuka, Bunkyo-ku, Tokyo 112-8610, Japan; 2The Glycoscience Institute, Ochanomizu University, 2-1-1 Otsuka, Bunkyo-ku, Tokyo 112-8610, Japan; and 3Laboratory of Cellular Biochemistry, RIKEN, 2-1 Hirosawa, Wako-shi, Saitama 351-0198, Japan
*To whom correspondence should be addressed. Tel: +81-3-5978-5345; Fax: +81-3-5978-5345; E-mail: kyoko{at}cc.ocha.ac.jp
Received November 8, 2006; Accepted November 13, 2006
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Abstract |
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Mammalian annexins are implicated in several physiological mechanisms based on their calcium-dependent phospholipid/membrane binding and carbohydrate-binding activities. In this study, we investigated gene expression profiles of all four Caenorhabditis elegans annexins, nex-1, -2, -3 and -4, throughout the development, and compared phospholipid- and carbohydrate-binding properties of their protein products, NEX-1, -2, -3 and -4. We found that nex-1 and -3 are transcribed continuously during the developmental stages, while expression of nex-2 and -4 appeared to be temporal, peaking at the L1 stage followed by a gradual decrease toward the adult stage. NEX-1 and -3 were detected as single protein band in total worm extracts by immunoblotting, but NEX-2 was heterogenic in size. NEX-1, -2, and -3 showed the binding activities to phosphatidylserine, phosphatidylinositol and phosphatidylethanolamine, but not to phosphatidylcholine. In contrast to their uniform phospholipids-binding properties, their glycosaminoglycan-binding activities were distinctive. NEX-2 bound to heparan sulfate and chondroitin, NEX-3 bound only to heparan sulfate, and NEX-1 showed no lectin activities under tested conditions. NEX-4 had neither phospholipids- nor carbohydrate-binding properties. Differentiated expression profiles and ligand-binding properties of NEX-1, -2, -3 and -4, shown in our study, may represent distinctive roles for each C. elegans annexins.
Key Words: annexin, lectin, C. elegans, glycosaminoglycan, phospholipid
Abbreviations: GST, glutathione S-transferase; GSH, glutathione; CBB, Coomassie Brilliant Blue; PVDF, polyvinylidene difluoride; HRP, horseradish peroxidase; FPLC, fast protein liquid chromatography; TBS, Tris-bufferd saline; GAG, glycosaminolycan; PC, phosphatidylchorine; PS, phosphatidylserine; PI, phosphatidylinositol; PE, phosphatidylethanolamine; POPC, palmitoyl-oleoyl-phosphatidylcholine; POPE, palmitoyl-oleoyl-phosphatidylethanolamine; POPS, palmitoyl-oleoyl-phosphatidylserine
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