%0 Journal Article %1 WOS:000374661800003 %A de Freitas, Cleverson D T %A da Cruz, Wallace T %A Silva, Maria Z R %A Vasconcelos, Ilka M %A Moreno, Frederico B M B %A Moreira, Renato A %A Monteiro-Moreira, Ana C O %A Alencar, Luciana M R %A Sousa, Jeanlex S %A Rocha, Bruno A M %A Ramos, Marcio V %C 233 SPRING ST, NEW YORK, NY 10013 USA %D 2016 %I SPRINGER %J PLANTA %K Apocynaceae; Peptidase; Proteome} force microscopy; {Atomic %N 5 %P 1115-1128 %R 10.1007/s00425-016-2468-8 %T Proteomic analysis and purification of an unusual germin-like protein with proteolytic activity in the latex of Thevetia peruviana %V 243 %X The latex from Thevetia peruviana is rich in plant defense proteins, including a 120 kDa cysteine peptidase with structural characteristics similar to germin-like proteins. More than 20,000 plant species produce latex, including Apocynaceae, Sapotaceae, Papaveraceae and Euphorbiaceae. To better understand the physiological role played by latex fluids, a proteomic analysis of Thevetia peruviana (Pers.) Schum latex was performed using two-dimensional gel electrophoresis and mass spectrometry. A total of 33 proteins (86 %) were identified, including storage proteins, a peptidase inhibitor, cysteine peptidases, peroxidases and osmotins. An unusual cysteine peptidase, termed peruvianin-I, was purified from the latex by a single chromatographic step involving gel filtration. The enzyme (glycoprotein) was inhibited by E-64 and iodoacetamide and exhibited high specific activity towards azocasein (K (m) 17.6 A mu M), with an optimal pH and temperature of 5.0-6.0 and 25-37 A degrees C, respectively. Gel filtration chromatography, two-dimensional gel electrophoresis, and mass spectrometry revealed that peruvianin-I possesses 120 kDa, pI 4.0, and six subunits (20 kDa). A unique N-terminal amino acid sequence was obtained to oligomer and monomers of peruvianin-I ((1)ADPGPLQDFCLADLNSPLFINGYPCRNPALAISDDF(36)). High-resolution images from atomic force microscopy showed the homohexameric structure of peruvianin-I may be organized as a trimer of dimers that form a central channel similar to germin-like proteins. Peruvianin-I exhibited no oxalate oxidase and superoxide dismutase activity or antifungal effects. Peruvianin-I represents the first germin-like protein (GLP) with cysteine peptidase activity, an activity unknown in the GLP family so far.

@article{WOS:000374661800003, abstract = {The latex from Thevetia peruviana is rich in plant defense proteins, including a 120 kDa cysteine peptidase with structural characteristics similar to germin-like proteins. More than 20,000 plant species produce latex, including Apocynaceae, Sapotaceae, Papaveraceae and Euphorbiaceae. To better understand the physiological role played by latex fluids, a proteomic analysis of Thevetia peruviana (Pers.) Schum latex was performed using two-dimensional gel electrophoresis and mass spectrometry. A total of 33 proteins (86 %) were identified, including storage proteins, a peptidase inhibitor, cysteine peptidases, peroxidases and osmotins. An unusual cysteine peptidase, termed peruvianin-I, was purified from the latex by a single chromatographic step involving gel filtration. The enzyme (glycoprotein) was inhibited by E-64 and iodoacetamide and exhibited high specific activity towards azocasein (K (m) 17.6 A mu M), with an optimal pH and temperature of 5.0-6.0 and 25-37 A degrees C, respectively. Gel filtration chromatography, two-dimensional gel electrophoresis, and mass spectrometry revealed that peruvianin-I possesses 120 kDa, pI 4.0, and six subunits (20 kDa). A unique N-terminal amino acid sequence was obtained to oligomer and monomers of peruvianin-I ((1)ADPGPLQDFCLADLNSPLFINGYPCRNPALAISDDF(36)). High-resolution images from atomic force microscopy showed the homohexameric structure of peruvianin-I may be organized as a trimer of dimers that form a central channel similar to germin-like proteins. Peruvianin-I exhibited no oxalate oxidase and superoxide dismutase activity or antifungal effects. Peruvianin-I represents the first germin-like protein (GLP) with cysteine peptidase activity, an activity unknown in the GLP family so far.}, added-at = {2022-05-23T20:00:14.000+0200}, address = {233 SPRING ST, NEW YORK, NY 10013 USA}, author = {de Freitas, Cleverson D T and da Cruz, Wallace T and Silva, Maria Z R and Vasconcelos, Ilka M and Moreno, Frederico B M B and Moreira, Renato A and Monteiro-Moreira, Ana C O and Alencar, Luciana M R and Sousa, Jeanlex S and Rocha, Bruno A M and Ramos, Marcio V}, biburl = {https://www.bibsonomy.org/bibtex/2257c4b45f0d99c1404859c1f0b801e0f/ppgfis_ufc_br}, doi = {10.1007/s00425-016-2468-8}, interhash = {8541362f25398cad086e38714880d6f7}, intrahash = {257c4b45f0d99c1404859c1f0b801e0f}, issn = {0032-0935}, journal = {PLANTA}, keywords = {Apocynaceae; Peptidase; Proteome} force microscopy; {Atomic}, number = 5, pages = {1115-1128}, publisher = {SPRINGER}, pubstate = {published}, timestamp = {2022-05-23T20:00:14.000+0200}, title = {Proteomic analysis and purification of an unusual germin-like protein with proteolytic activity in the latex of Thevetia peruviana}, tppubtype = {article}, volume = 243, year = 2016 }