Crystal structure of cardosin A, a glycosylated and Arg-Gly-Asp-containing aspartic proteinase from the flowers of Cynara cardunculus L.
Aspartic proteinases (AP) have been widely studied within the living world, but so far no plant AP have been structurally characterized. The refined cardosin A crystallographic structure includes two molecules, built up by two glycosylated peptide chains (31 and 15 kDa each). The fold of cardosin A is typical within the AP family. The glycosyl content is described by 19 sugar rings attached to Asn-67 and Asn-257. They are localized on the molecular surface away from the conserved active site and show a new glycan of the plant complex type. A hydrogen bond between Gln-126 and Manbeta4 renders the monosaccharide oxygen O-2 sterically inaccessible to accept a xylosyl residue, therefore explaining the new type of the identified plant glycan. The Arg-Gly-Asp sequence, which has been shown to be involved in recognition of a putative cardosin A receptor, was found in a loop between two beta-strands on the molecular surface opposite the active site cleft. Based on the crystal structure, a possible mechanism whereby cardosin A might be orientated at the cell surface of the style to interact with its putative receptor from pollen is proposed. The biological implications of these findings are also discussed.
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Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Apartado 127, 2780-Oeiras, Portugal.
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Costa
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Soares
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Veríssimo
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Pires
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Cooper
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D000595:Amino Acid Sequence; D000818:Animals; D001216:Asparagine; D016282:Aspartic Acid Endopeptidases / Q000737:chemistry* / Q000302:isolation & purification; D001665:Binding Sites; D018360:Crystallography, X-Ray; D006031:Glycosylation; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000737:chemistry* / Q000302:isolation & purification; D018547:Plant Stems / Q000201:enzymology; D010944:Plants / Q000201:enzymology*; D017433:Protein Structure, Secondary
D010940:Plant Proteins; D001216:Asparagine; D016282:Aspartic Acid Endopeptidases; C097455:aspartic proteinases, Cynara cardunculus