tailieunhanh - Báo cáo khoa học: Molecular analysis of the interaction between cardosin A and phospholipase Da Identification of RGD/KGE sequences as binding motifs for C2 domains

Cardosin A is an RGD-containing aspartic proteinase from the stigmatic papillae ofCynara cardunculusL. A putative cardosin A-binding protein has previously been isolated from pollen suggesting its potential involve-ment in pollen–pistil interaction [Faro C, Ramalho-Santos M, Vieira M, Mendes A, Simo˜es I, Andrade R, Verissimo P, Lin X, Tang J & Pires E (1999) J Biol Chem274, 28724–28729]. | iFEBS Journal Molecular analysis of the interaction between cardosin A and phospholipase Da Identification of RGD KGE sequences as binding motifs for C2 domains Isaura Simoes1 Eva-Christina Mueller2 Albrecht Otto2 Daniel Bur3 Alice Y. Cheung4 Carlos Faro1 and Euclides Pires1 1 Departamento de Biologia Molecular e Biotecnologia Centro de Neurociencias e Biologia Celular Universidade de Coimbra and Departamento de Bioquimica Faculdade de Ciencias e Tecnologia Universidade de Coimbra Portugal 2 Max Delbrueck Center for Molecular Medicine Berlin Germany 3 Actelion Pharmaceuticals Ltd Allschwil Switzerland 4 Department of Biochemistry and Molecular Biology University of Massachusetts Amherst MA USA Keywords aspartic proteinases C2 domain cardosin A phospholipase D RGD KGE sequences Correspondence C. Faro Departamento de Bioquimica Universidade de Coimbra Apt. 3126 3000 Coimbra Portugal Fax 351 239 480208 Tel 351 239 480210 E-mail cfaro@ Note The nucleotide sequence of PLDa from C. cardunculus L has been submitted to the EBI Data Bank with the accession number AJ583515 Received 9 June 2005 revised 27 July 2005 accepted 14 September 2005 doi 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 Simoes 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 Da 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-trans-ferase-fused phospholipase Da constructs were performed. Results revealed that the C2 domain .