Identification and molecular analysis of interaction sites in the MtSEO-F1 protein involved in forisome assembly
| Autor: | Richard M. Twyman, Judith Rose, Boje Müller, Matthias Senft, Sira Groscurth, Gundula A. Noll, Franziska Visser, Dirk Prüfer, Kevin F. Sicking |
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| Rok vydání: | 2020 |
| Předmět: |
Protein Conformation
Sequence (biology) 02 engineering and technology Thioredoxin fold Biochemistry Hydrophobic effect 03 medical and health sciences Bimolecular fluorescence complementation Forisome Gene Expression Regulation Plant Structural Biology Amino Acid Sequence Molecular Biology Gene Library Plant Proteins 030304 developmental biology chemistry.chemical_classification 0303 health sciences biology Plant Extracts Mutagenesis Fabaceae General Medicine Plants Genetically Modified 021001 nanoscience & nanotechnology biology.organism_classification Medicago truncatula Amino acid chemistry Biophysics Protein Multimerization 0210 nano-technology Hydrophobic and Hydrophilic Interactions |
| Zdroj: | International Journal of Biological Macromolecules. 144:603-614 |
| ISSN: | 0141-8130 |
| DOI: | 10.1016/j.ijbiomac.2019.12.092 |
| Popis: | Forisomes are large mechanoprotein complexes found solely in legumes such as Medicago truncatula. They comprise several "sieve element occlusion by forisome" (SEO-F) subunits, with MtSEO-F1 as the major structure-forming component. SEO-F proteins possess three conserved domains -an N-terminal domain (SEO-NTD), a potential thioredoxin fold, and a C-terminal domain (SEO-CTD)- but structural and biochemical data are scarce and little is known about the contribution of these domains to forisome assembly. To identify key amino acids involved in MtSEO-F1 dimerization and complex formation, we investigated protein-protein interactions by bimolecular fluorescence complementation and the analysis of yeast two-hybrid and random mutagenesis libraries. We identified a SEO-NTD core region as the major dimerization site, with abundant hydrophobic residues and rare charged residues suggesting dimerization is driven by the hydrophobic effect. We also found that ~45% of the full-length MtSEO-F1 sequence must be conserved for higher-order protein assembly, indicating that large interaction surfaces facilitate stable interactions, contributing to the high resilience of forisome bodies. Interestingly, the removal of 62 amino acids from the C-terminus did not disrupt forisome assembly. This is the first study unraveling interaction sites and mechanisms within the MtSEO-F1 protein at the level of dimerization and complex formation. |
| Databáze: | OpenAIRE |
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