Ligand-binding properties of annexin from Caenorhabditis elegans (annexin XVI, Nex-1).
Annexins are structurally related proteins that bind phospholipids in a calcium-dependent manner. Recently, we showed that annexins IV, V, and VI also bind glycosaminoglycans in a calcium-dependent manner. Annexins are widely distributed from lower to higher eukaryotes, and the nematode Caenorhabditis elegans has been found to contain Nex-1, an annexin homologue. Here, we characterize the ligand-binding properties of Nex-1 using recombinant Nex-1. Nex-1 binds to liposomes containing phosphatidylserine. The apparent K(d) was calculated by Biacore to be 4.4 nM. Compared to mammalian annexins, the Nex-1 phospholipid-binding specificities were similar whereas the K(d) values were one order of magnitude larger. The Nex-1 glycosaminoglycan-binding specificities were investigated by affinity chromatography and solid-phase assays. Nex-1 binds to heparin, heparan sulfate, and chondroitin sulfate but not to chondroitin and chemically N- or O-desulfated heparin. Besides phospholipids, heparan sulfate and/or chondroitin (sulfate), probably on perlecan, could be endogenous ligands of Nex-1.
Satoh
A
A
The Graduate School of Humanities and Sciences, Department of Chemistry, Faculty of Science, Ochanomizu University, Otsuka Bunkyo-ku, Tokyo 112-8610, Japan. isamu@cc.ocha.ac.jp
Miwa
H E
HE
Kojima
K
K
Hirabayashi
J
J
Matsumoto
I
I
Hazuki
M
M
D000818:Animals; D017302:Annexins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D001667:Binding, Competitive; D017173:Caenorhabditis elegans / Q000737:chemistry*; D029742:Caenorhabditis elegans Proteins; D002809:Chondroitin Sulfates / Q000378:metabolism; D002846:Chromatography, Affinity; D017931:DNA Primers / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D005822:Genetic Vectors; D006025:Glycosaminoglycans / Q000378:metabolism*; D015801:Helminth Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D006493:Heparin / Q000378:metabolism; D006497:Heparan Sulfate / Q000378:metabolism; D008024:Ligands; D008081:Liposomes / Q000737:chemistry; D010718:Phosphatidylserines / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D011485:Protein Binding; D011994:Recombinant Proteins / Q000378:metabolism
D017302:Annexins; D029742:Caenorhabditis elegans Proteins; D002809:Chondroitin Sulfates; D017931:DNA Primers; D006025:Glycosaminoglycans; D015801:Helminth Proteins; D006493:Heparin; D006497:Heparan Sulfate; D008024:Ligands; D008081:Liposomes; D010718:Phosphatidylserines; D011994:Recombinant Proteins; C098890:Nex-1 protein, C elegans