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Molecular analysis of the interaction between cardosin A and phospholipase Dalpha

Item Type:Article
Title:Molecular analysis of the interaction between cardosin A and phospholipase Dalpha
Creators Name:Simoes, I. and Mueller, E.C. and Otto, A. and Bur, D. and Cheung, A.Y. and Faro, C. and Pires, E.
Abstract:Cardosin A is an RGD-containing aspartic proteinase from the stigmatic papillae of Cynara cardunculus L. A putative cardosin A-binding protein has previously been isolated from pollen suggesting its potential involvement in pollen-pistil interaction [Faro C, Ramalho-Santos M, Vieira M, Mendes A, Simoees I, Andrade R, Verissimo P, Lin X, Tang J & Pires E (1999) J Biol Chem 274, 28724-28729]. Here we report the identification of phospholipase Dα as a cardosin A-binding protein. The interaction was confirmed by coimmunoprecipitation studies and pull-down assays. To investigate the structural and molecular determinants involved in the interaction, pull-down assays with cardosin A and various glutathione S-transferase-fused phospholipase D{alpha} constructs were performed. Results revealed that the C2 domain of phospholipase D{alpha} contains the cardosin A-binding activity. Further assays with mutated recombinant forms of cardosin A showed that the RGD motif as well as the unprecedented KGE motif, which is structurally and charge-wise very similar to RGD, are indispensable for the interaction. Taken together our results indicate that the C2 domain of plant phospholipase Dα can act as a cardosin A-binding domain and suggest that plant C2 domains may have an additional role as RGD/KGE-recognition domains.
Keywords:Aspartic Proteinases, C2 Domain, Cardosin A, Phospholipase D, RGD/KGE Sequences
Source:FEBS Journal
Publisher:Blackwell Publishing
Page Range:5786-5798
Date:1 November 2005
Official Publication:https://doi.org/10.1111/j.1742-4658.2005.04967.x
PubMed:View item in PubMed

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