Proteomic and Targeted Lipidomic Analyses of Fluid and Rigid Rubber Particle Membrane Domains in Guayule
Rubber (<i>cis</i>-1,4-polyisoprene) is produced in cytosolic unilamellar vesicles called rubber particles (RPs), and the protein complex responsible for this synthesis, the rubber transferase (RTase), is embedded in, or tethered to, the membranes of these RPs. Solubilized enzyme activit...
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2024-10-01
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| author | Joshua J. Blakeslee Eun-Hyang Han Yun Lin Jinshan Lin Seema Nath Liwen Zhang Zhenyu Li Katrina Cornish |
| author_facet | Joshua J. Blakeslee Eun-Hyang Han Yun Lin Jinshan Lin Seema Nath Liwen Zhang Zhenyu Li Katrina Cornish |
| author_sort | Joshua J. Blakeslee |
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| description | Rubber (<i>cis</i>-1,4-polyisoprene) is produced in cytosolic unilamellar vesicles called rubber particles (RPs), and the protein complex responsible for this synthesis, the rubber transferase (RTase), is embedded in, or tethered to, the membranes of these RPs. Solubilized enzyme activity is very difficult to achieve because the polymerization of highly hydrophilic substrates into hydrophobic polymers requires a polar/non-polar interface and a hydrophobic compartment. Using guayule (<i>Parthenium argentatum</i>) as a model rubber-producing species, we optimized methods to isolate RP unilamellear membranes and then a subset of membrane microdomains (detergent-resistant membranes) likely to contain protein complexes such as RTase. The phospholipid and sterol composition of these membranes and microdomains were analyzed using thin-layer chromatography (TLC) and liquid chromatography tandem mass spectroscopy (LC-MS/MS). Our data indicate that RP membranes consist predominantly of phosphatidic acid-containing membrane microdomains (DRMs or “lipid rafts”). Proteomic analyses of guayule RP membranes and membrane microdomains identified 80 putative membrane proteins covering 30 functional categories. From this population, we have tentatively identified several proteins in multiple functional domains associated with membrane microdomains which may be critical to RTase function. Definition of the mechanisms underlying rubber synthesis will provide targets for both metabolic engineering and breeding strategies designed to increase natural rubber production in latex-producing species. |
| format | Article |
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| institution | OA Journals |
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| language | English |
| publishDate | 2024-10-01 |
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| spelling | doaj-art-ec6dd825337449d29a0602fb4807d2c02025-08-20T02:13:18ZengMDPI AGPlants2223-77472024-10-011321297010.3390/plants13212970Proteomic and Targeted Lipidomic Analyses of Fluid and Rigid Rubber Particle Membrane Domains in GuayuleJoshua J. Blakeslee0Eun-Hyang Han1Yun Lin2Jinshan Lin3Seema Nath4Liwen Zhang5Zhenyu Li6Katrina Cornish7Department of Horticulture and Crop Science, Ohio Agricultural Research and Development Center (OARDC), The Ohio State University, Wooster, OH 44691, USADepartment of Horticulture and Crop Science, Ohio Agricultural Research and Development Center (OARDC), The Ohio State University, Wooster, OH 44691, USADepartment of Horticulture and Crop Science, Ohio Agricultural Research and Development Center (OARDC), The Ohio State University, Wooster, OH 44691, USALaboratory for the Analysis of Metabolites from Plants (LAMP), The Ohio State University, Columbus, OH 43210, USALaboratory for the Analysis of Metabolites from Plants (LAMP), The Ohio State University, Columbus, OH 43210, USACampus Chemical Instrumentation Center (CCIC), The Ohio State University, Columbus, OH 43210, USADepartment of Horticulture and Crop Science, Ohio Agricultural Research and Development Center (OARDC), The Ohio State University, Wooster, OH 44691, USADepartment of Horticulture and Crop Science, Ohio Agricultural Research and Development Center (OARDC), The Ohio State University, Wooster, OH 44691, USARubber (<i>cis</i>-1,4-polyisoprene) is produced in cytosolic unilamellar vesicles called rubber particles (RPs), and the protein complex responsible for this synthesis, the rubber transferase (RTase), is embedded in, or tethered to, the membranes of these RPs. Solubilized enzyme activity is very difficult to achieve because the polymerization of highly hydrophilic substrates into hydrophobic polymers requires a polar/non-polar interface and a hydrophobic compartment. Using guayule (<i>Parthenium argentatum</i>) as a model rubber-producing species, we optimized methods to isolate RP unilamellear membranes and then a subset of membrane microdomains (detergent-resistant membranes) likely to contain protein complexes such as RTase. The phospholipid and sterol composition of these membranes and microdomains were analyzed using thin-layer chromatography (TLC) and liquid chromatography tandem mass spectroscopy (LC-MS/MS). Our data indicate that RP membranes consist predominantly of phosphatidic acid-containing membrane microdomains (DRMs or “lipid rafts”). Proteomic analyses of guayule RP membranes and membrane microdomains identified 80 putative membrane proteins covering 30 functional categories. From this population, we have tentatively identified several proteins in multiple functional domains associated with membrane microdomains which may be critical to RTase function. Definition of the mechanisms underlying rubber synthesis will provide targets for both metabolic engineering and breeding strategies designed to increase natural rubber production in latex-producing species.https://www.mdpi.com/2223-7747/13/21/2970guayulemembranelipidomicsproteomicsrubber particle membranesrubber transferase complex |
| spellingShingle | Joshua J. Blakeslee Eun-Hyang Han Yun Lin Jinshan Lin Seema Nath Liwen Zhang Zhenyu Li Katrina Cornish Proteomic and Targeted Lipidomic Analyses of Fluid and Rigid Rubber Particle Membrane Domains in Guayule Plants guayule membrane lipidomics proteomics rubber particle membranes rubber transferase complex |
| title | Proteomic and Targeted Lipidomic Analyses of Fluid and Rigid Rubber Particle Membrane Domains in Guayule |
| title_full | Proteomic and Targeted Lipidomic Analyses of Fluid and Rigid Rubber Particle Membrane Domains in Guayule |
| title_fullStr | Proteomic and Targeted Lipidomic Analyses of Fluid and Rigid Rubber Particle Membrane Domains in Guayule |
| title_full_unstemmed | Proteomic and Targeted Lipidomic Analyses of Fluid and Rigid Rubber Particle Membrane Domains in Guayule |
| title_short | Proteomic and Targeted Lipidomic Analyses of Fluid and Rigid Rubber Particle Membrane Domains in Guayule |
| title_sort | proteomic and targeted lipidomic analyses of fluid and rigid rubber particle membrane domains in guayule |
| topic | guayule membrane lipidomics proteomics rubber particle membranes rubber transferase complex |
| url | https://www.mdpi.com/2223-7747/13/21/2970 |
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