Enrichment of Phosphoproteins and Phosphopeptide Derivatization Identify Universal Stress Proteins in Elicitor-Treated Arabidopsis
Protein phosphorylation is a key biological process that regulates reactions involved in plant-microbe interactions. The phosphorylated form of a protein often represents only a small fraction of the total population and can be problematic to analyze in a mass spectrometer. We demonstrate how a tita...
| Main Authors: | , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
The American Phytopathological Society
2008-10-01
|
| Series: | Molecular Plant-Microbe Interactions |
| Subjects: | |
| Online Access: | https://apsjournals.apsnet.org/doi/10.1094/MPMI-21-10-1275 |
| _version_ | 1819085191238385664 |
|---|---|
| author | Marit Lenman Carolin Sörensson Erik Andreasson |
| author_facet | Marit Lenman Carolin Sörensson Erik Andreasson |
| author_sort | Marit Lenman |
| collection | DOAJ |
| description | Protein phosphorylation is a key biological process that regulates reactions involved in plant-microbe interactions. The phosphorylated form of a protein often represents only a small fraction of the total population and can be problematic to analyze in a mass spectrometer. We demonstrate how a titanium dioxide (TiO2) resin can be employed for the enrichment of phosphoproteins, as well as a method to derivatize TiO2-purified phosphopeptides to facilitate determination of the exact site of phosphorylation. The use of these methods was exemplified by the identification of two plant proteins that were shown to be phosphorylated after the elicitation of Arabidopsis cells with Phytophthora infestans zoospores and xylanase. Both of the proteins that were identified, At5g54430.1 and At4g27320.1, were found to contain a universal stress protein domain with conserved residues for ATP binding. |
| first_indexed | 2024-12-21T21:00:26Z |
| format | Article |
| id | doaj.art-9b4a04f1cf434708a69b86e740750e1b |
| institution | Directory Open Access Journal |
| issn | 0894-0282 1943-7706 |
| language | English |
| last_indexed | 2024-12-21T21:00:26Z |
| publishDate | 2008-10-01 |
| publisher | The American Phytopathological Society |
| record_format | Article |
| series | Molecular Plant-Microbe Interactions |
| spelling | doaj.art-9b4a04f1cf434708a69b86e740750e1b2022-12-21T18:50:27ZengThe American Phytopathological SocietyMolecular Plant-Microbe Interactions0894-02821943-77062008-10-0121101275128410.1094/MPMI-21-10-1275Enrichment of Phosphoproteins and Phosphopeptide Derivatization Identify Universal Stress Proteins in Elicitor-Treated ArabidopsisMarit LenmanCarolin SörenssonErik AndreassonProtein phosphorylation is a key biological process that regulates reactions involved in plant-microbe interactions. The phosphorylated form of a protein often represents only a small fraction of the total population and can be problematic to analyze in a mass spectrometer. We demonstrate how a titanium dioxide (TiO2) resin can be employed for the enrichment of phosphoproteins, as well as a method to derivatize TiO2-purified phosphopeptides to facilitate determination of the exact site of phosphorylation. The use of these methods was exemplified by the identification of two plant proteins that were shown to be phosphorylated after the elicitation of Arabidopsis cells with Phytophthora infestans zoospores and xylanase. Both of the proteins that were identified, At5g54430.1 and At4g27320.1, were found to contain a universal stress protein domain with conserved residues for ATP binding.https://apsjournals.apsnet.org/doi/10.1094/MPMI-21-10-12754-sulfophenyl isothiocyanateSPITC |
| spellingShingle | Marit Lenman Carolin Sörensson Erik Andreasson Enrichment of Phosphoproteins and Phosphopeptide Derivatization Identify Universal Stress Proteins in Elicitor-Treated Arabidopsis Molecular Plant-Microbe Interactions 4-sulfophenyl isothiocyanate SPITC |
| title | Enrichment of Phosphoproteins and Phosphopeptide Derivatization Identify Universal Stress Proteins in Elicitor-Treated Arabidopsis |
| title_full | Enrichment of Phosphoproteins and Phosphopeptide Derivatization Identify Universal Stress Proteins in Elicitor-Treated Arabidopsis |
| title_fullStr | Enrichment of Phosphoproteins and Phosphopeptide Derivatization Identify Universal Stress Proteins in Elicitor-Treated Arabidopsis |
| title_full_unstemmed | Enrichment of Phosphoproteins and Phosphopeptide Derivatization Identify Universal Stress Proteins in Elicitor-Treated Arabidopsis |
| title_short | Enrichment of Phosphoproteins and Phosphopeptide Derivatization Identify Universal Stress Proteins in Elicitor-Treated Arabidopsis |
| title_sort | enrichment of phosphoproteins and phosphopeptide derivatization identify universal stress proteins in elicitor treated arabidopsis |
| topic | 4-sulfophenyl isothiocyanate SPITC |
| url | https://apsjournals.apsnet.org/doi/10.1094/MPMI-21-10-1275 |
| work_keys_str_mv | AT maritlenman enrichmentofphosphoproteinsandphosphopeptidederivatizationidentifyuniversalstressproteinsinelicitortreatedarabidopsis AT carolinsorensson enrichmentofphosphoproteinsandphosphopeptidederivatizationidentifyuniversalstressproteinsinelicitortreatedarabidopsis AT erikandreasson enrichmentofphosphoproteinsandphosphopeptidederivatizationidentifyuniversalstressproteinsinelicitortreatedarabidopsis |