Leucine-rich-repeat-containing variable lymphocyte receptors as modules to target plant-expressed proteins
Abstract Background The ability to target and manipulate protein-based cellular processes would accelerate plant research; yet, the technology to specifically and selectively target plant-expressed proteins is still in its infancy. Leucine-rich repeats (LRRs) are ubiquitously present protein domains...
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| Format: | Article |
| Language: | English |
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BMC
2017-04-01
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| Series: | Plant Methods |
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| Online Access: | http://link.springer.com/article/10.1186/s13007-017-0180-8 |
| _version_ | 1818266534479396864 |
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| author | André C. Velásquez Kinya Nomura Max D. Cooper Brantley R. Herrin Sheng Yang He |
| author_facet | André C. Velásquez Kinya Nomura Max D. Cooper Brantley R. Herrin Sheng Yang He |
| author_sort | André C. Velásquez |
| collection | DOAJ |
| description | Abstract Background The ability to target and manipulate protein-based cellular processes would accelerate plant research; yet, the technology to specifically and selectively target plant-expressed proteins is still in its infancy. Leucine-rich repeats (LRRs) are ubiquitously present protein domains involved in mediating protein–protein interactions. LRRs confer the binding specificity to the highly diverse variable lymphocyte receptor (VLR) antibodies (including VLRA, VLRB and VLRC types) that jawless vertebrates make as the functional equivalents of jawed vertebrate immunoglobulin-based antibodies. Results In this study, VLRBs targeting an effector protein from a plant pathogen, HopM1, were developed by immunizing lampreys and using yeast surface display to select for high-affinity VLRBs. HopM1-specific VLRBs (VLRM1) were expressed in planta in the cytosol, the trans-Golgi network, and the apoplast. Expression of VLRM1 was higher when the protein localized to an oxidizing environment that would favor disulfide bridge formation (when VLRM1 was not localized to the cytoplasm), as disulfide bonds are necessary for proper VLR folding. VLRM1 specifically interacted in planta with HopM1 but not with an unrelated bacterial effector protein while HopM1 failed to interact with a non-specific VLRB. Conclusions In the future, VLRs may be used as flexible modules to bind proteins or carbohydrates of interest in planta, with broad possibilities for their use by binding directly to their targets and inhibiting their action, or by creating chimeric proteins with new specificities in which endogenous LRR domains are replaced by those present in VLRs. |
| first_indexed | 2024-12-12T20:08:14Z |
| format | Article |
| id | doaj.art-277cd41654de4fe2927cbc8fafc469aa |
| institution | Directory Open Access Journal |
| issn | 1746-4811 |
| language | English |
| last_indexed | 2024-12-12T20:08:14Z |
| publishDate | 2017-04-01 |
| publisher | BMC |
| record_format | Article |
| series | Plant Methods |
| spelling | doaj.art-277cd41654de4fe2927cbc8fafc469aa2022-12-22T00:13:35ZengBMCPlant Methods1746-48112017-04-0113111610.1186/s13007-017-0180-8Leucine-rich-repeat-containing variable lymphocyte receptors as modules to target plant-expressed proteinsAndré C. Velásquez0Kinya Nomura1Max D. Cooper2Brantley R. Herrin3Sheng Yang He4DOE Plant Research Laboratory, Michigan State UniversityDOE Plant Research Laboratory, Michigan State UniversityDepartment of Pathology and Laboratory Medicine, Emory UniversityDepartment of Pathology and Laboratory Medicine, Emory UniversityDOE Plant Research Laboratory, Michigan State UniversityAbstract Background The ability to target and manipulate protein-based cellular processes would accelerate plant research; yet, the technology to specifically and selectively target plant-expressed proteins is still in its infancy. Leucine-rich repeats (LRRs) are ubiquitously present protein domains involved in mediating protein–protein interactions. LRRs confer the binding specificity to the highly diverse variable lymphocyte receptor (VLR) antibodies (including VLRA, VLRB and VLRC types) that jawless vertebrates make as the functional equivalents of jawed vertebrate immunoglobulin-based antibodies. Results In this study, VLRBs targeting an effector protein from a plant pathogen, HopM1, were developed by immunizing lampreys and using yeast surface display to select for high-affinity VLRBs. HopM1-specific VLRBs (VLRM1) were expressed in planta in the cytosol, the trans-Golgi network, and the apoplast. Expression of VLRM1 was higher when the protein localized to an oxidizing environment that would favor disulfide bridge formation (when VLRM1 was not localized to the cytoplasm), as disulfide bonds are necessary for proper VLR folding. VLRM1 specifically interacted in planta with HopM1 but not with an unrelated bacterial effector protein while HopM1 failed to interact with a non-specific VLRB. Conclusions In the future, VLRs may be used as flexible modules to bind proteins or carbohydrates of interest in planta, with broad possibilities for their use by binding directly to their targets and inhibiting their action, or by creating chimeric proteins with new specificities in which endogenous LRR domains are replaced by those present in VLRs.http://link.springer.com/article/10.1186/s13007-017-0180-8Protein targetingLeucine-rich repeatVariable lymphocyte receptorModulesHopM1 |
| spellingShingle | André C. Velásquez Kinya Nomura Max D. Cooper Brantley R. Herrin Sheng Yang He Leucine-rich-repeat-containing variable lymphocyte receptors as modules to target plant-expressed proteins Plant Methods Protein targeting Leucine-rich repeat Variable lymphocyte receptor Modules HopM1 |
| title | Leucine-rich-repeat-containing variable lymphocyte receptors as modules to target plant-expressed proteins |
| title_full | Leucine-rich-repeat-containing variable lymphocyte receptors as modules to target plant-expressed proteins |
| title_fullStr | Leucine-rich-repeat-containing variable lymphocyte receptors as modules to target plant-expressed proteins |
| title_full_unstemmed | Leucine-rich-repeat-containing variable lymphocyte receptors as modules to target plant-expressed proteins |
| title_short | Leucine-rich-repeat-containing variable lymphocyte receptors as modules to target plant-expressed proteins |
| title_sort | leucine rich repeat containing variable lymphocyte receptors as modules to target plant expressed proteins |
| topic | Protein targeting Leucine-rich repeat Variable lymphocyte receptor Modules HopM1 |
| url | http://link.springer.com/article/10.1186/s13007-017-0180-8 |
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