FRINK Linked Data Fragments server

Matches in SemOpenAlex for { <https://semopenalex.org/work/W3082023227> ?p ?o ?g. }

• W3082023227 abstract "Up-to-now, several biochemical methods have been developed to allow specific organelle isolation from plant tissues. These procedures are often time consuming, require substantial amounts of plant material, have low yield or do not result in pure organelle fractions. Moreover, barely a protocol allows rapid and flexible isolation of different subcellular compartments. The recently published SpySystem enables the in vitro and in vivo covalent linkage between proteins and protein complexes. Here we describe the use of this system to tag and purify plant organelles.We developed a simple and specific method to in vivo tag and visualize, as well as isolate organelles of interest from crude plant extracts. This was achieved by expressing the covalent split-isopeptide interaction system, consisting of SpyTag and SpyCatcher, in Nicotiana benthamiana leaves. The functionality of the SpySystem in planta, combined with downstream applications, was proven. Using organelle-specific membrane anchor sequences to program the sub-cellular localization of the SpyTag peptide, we could tag the outer envelope of chloroplasts and mitochondria. By co-expression of a cytosolic, soluble eGFP-SpyCatcher fusion protein, we could demonstrate intermolecular isopeptide formation in planta and proper organelle targeting of the SpyTag peptides to the respective organelles. For one-step organelle purification, recombinantly expressed SpyCatcher protein was immobilized on magnetic microbeads via covalent thiol-etherification. To isolate tagged organelles, crude plant filtrates were mixed with SpyCatcher-coated beads which allowed isolation of SpyTag-labelled chloroplasts and mitochondria. The isolated organelles were intact, showed high yield and hardly contaminants and can be subsequently used for further molecular or biochemical analysis.The SpySystem can be used to in planta label subcellular structures, which enables the one-step purification of organelles from crude plant extracts. The beauty of the system is that it works as a covalent toolbox. Labeling of different organelles with individual tags under control of cell-specific and/or inducible promoter sequences will allow the rapid organelle and cell-type specific purification. Simultaneous labeling of different organelles with specific Tag/Catcher combinations will enable simultaneous isolation of different organelles from one plant extract in future experiments." @default.
• W3082023227 created "2020-09-08" @default.
• W3082023227 creator A5037812041 @default.
• W3082023227 creator A5054613791 @default.
• W3082023227 creator A5057644103 @default.
• W3082023227 creator A5086479317 @default.
• W3082023227 date "2020-09-01" @default.
• W3082023227 modified "2023-10-18" @default.
• W3082023227 title "Tagging and catching: rapid isolation and efficient labeling of organelles using the covalent Spy-System in planta" @default.
• W3082023227 cites W1511869459 @default.
• W3082023227 cites W16131576 @default.
• W3082023227 cites W1925510343 @default.
• W3082023227 cites W1965392584 @default.
• W3082023227 cites W1976672416 @default.
• W3082023227 cites W1990591017 @default.
• W3082023227 cites W1992672925 @default.
• W3082023227 cites W1994963086 @default.
• W3082023227 cites W1996192845 @default.
• W3082023227 cites W1996310347 @default.
• W3082023227 cites W2003842357 @default.
• W3082023227 cites W2014839507 @default.
• W3082023227 cites W2015815278 @default.
• W3082023227 cites W2016104233 @default.
• W3082023227 cites W2016841036 @default.
• W3082023227 cites W2029069158 @default.
• W3082023227 cites W2036156669 @default.
• W3082023227 cites W2038551788 @default.
• W3082023227 cites W2053716778 @default.
• W3082023227 cites W2061226620 @default.
• W3082023227 cites W2065372417 @default.
• W3082023227 cites W2069830527 @default.
• W3082023227 cites W2073862341 @default.
• W3082023227 cites W2094314820 @default.
• W3082023227 cites W2100738905 @default.
• W3082023227 cites W2108733083 @default.
• W3082023227 cites W2111198605 @default.
• W3082023227 cites W2117286856 @default.
• W3082023227 cites W2121115207 @default.
• W3082023227 cites W2121115364 @default.
• W3082023227 cites W2123838396 @default.
• W3082023227 cites W2124784707 @default.
• W3082023227 cites W2135194116 @default.
• W3082023227 cites W2151007289 @default.
• W3082023227 cites W2159490982 @default.
• W3082023227 cites W2169536239 @default.
• W3082023227 cites W2169581536 @default.
• W3082023227 cites W2174272296 @default.
• W3082023227 cites W2235815306 @default.
• W3082023227 cites W2263431558 @default.
• W3082023227 cites W2337081411 @default.
• W3082023227 cites W2346386833 @default.
• W3082023227 cites W2418667966 @default.
• W3082023227 cites W2440721038 @default.
• W3082023227 cites W2483356696 @default.
• W3082023227 cites W2517861725 @default.
• W3082023227 cites W253061803 @default.
• W3082023227 cites W2541529771 @default.
• W3082023227 cites W2555146968 @default.
• W3082023227 cites W2590803367 @default.
• W3082023227 cites W2607918806 @default.
• W3082023227 cites W2731380181 @default.
• W3082023227 cites W2767276910 @default.
• W3082023227 cites W2768553058 @default.
• W3082023227 cites W2799715258 @default.
• W3082023227 cites W2800937816 @default.
• W3082023227 cites W2811496274 @default.
• W3082023227 cites W2852019624 @default.
• W3082023227 cites W2898989665 @default.
• W3082023227 cites W2901897123 @default.
• W3082023227 cites W2943089648 @default.
• W3082023227 cites W2953108757 @default.
• W3082023227 cites W2966261754 @default.
• W3082023227 cites W2972369752 @default.
• W3082023227 cites W2991309876 @default.
• W3082023227 cites W2996724171 @default.
• W3082023227 cites W3000743194 @default.
• W3082023227 cites W3000906391 @default.
• W3082023227 cites W3006482621 @default.
• W3082023227 cites W4240402038 @default.
• W3082023227 cites W4250825251 @default.
• W3082023227 doi "https://doi.org/10.1186/s13007-020-00663-9" @default.
• W3082023227 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/7465787" @default.
• W3082023227 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/32905125" @default.
• W3082023227 hasPublicationYear "2020" @default.
• W3082023227 type Work @default.
• W3082023227 sameAs 3082023227 @default.
• W3082023227 citedByCount "4" @default.
• W3082023227 countsByYear W30820232272021 @default.
• W3082023227 countsByYear W30820232272023 @default.
• W3082023227 crossrefType "journal-article" @default.
• W3082023227 hasAuthorship W3082023227A5037812041 @default.
• W3082023227 hasAuthorship W3082023227A5054613791 @default.
• W3082023227 hasAuthorship W3082023227A5057644103 @default.
• W3082023227 hasAuthorship W3082023227A5086479317 @default.
• W3082023227 hasBestOaLocation W30820232271 @default.
• W3082023227 hasConcept C104317684 @default.
• W3082023227 hasConcept C168240541 @default.
• W3082023227 hasConcept C178790620 @default.
• W3082023227 hasConcept C180577832 @default.
• W3082023227 hasConcept C185592680 @default.

• next