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| Content Provider | Springer Nature Link |
|---|---|
| Author | Hagel, Jillian M. Facchini, Peter J. |
| Copyright Year | 2012 |
| Abstract | Benzylisoquinoline alkaloids are a large and structurally diverse group of natural plant products that includes many compounds with potent biological activities, including the antimicrobial agent sanguinarine. The putative subcellular localization of the sanguinarine pathway was determined using in-frame N-terminal fusions between cDNAs encoding nine consecutive biosynthetic enzymes and the gene encoding the green fluorescent protein (GFP). Expression constructs were introduced into cultured opium poppy cells by particle bombardment, and the localization of fusion proteins was visualized using epifluorescence microscopy. GFP fusions with two O-methyltransferases and two N-methyltransferases in the sanguinarine pathway all produced non-targeted fluorescence in the cytosol and nucleus. Interspersed between these soluble proteins are five membrane-bound cytochromes P450. Corresponding cDNAs are available for three P450s, all of which produced fluorescence when fused to GFP in association with the endoplasmic reticulum (ER). Two enzymes with suggested or known N-terminal signal peptides were initially associated with the ER, but were subsequently transported to the central vacuole suggesting their occurrence in the ER lumen. The alternating localization of these biosynthetic enzymes to three subcellular compartments indicates extensive trafficking of pathway intermediates across the endomembranes and suggests a key role for compartmentalization in the regulation of sanguinarine metabolism. |
| Starting Page | 233 |
| Ending Page | 240 |
| Page Count | 8 |
| File Format | |
| ISSN | 10545476 |
| Journal | In Vitro Cellular & Developmental Biology - Plant |
| Volume Number | 48 |
| Issue Number | 2 |
| e-ISSN | 14752689 |
| Language | English |
| Publisher | Springer-Verlag |
| Publisher Date | 2012-03-23 |
| Publisher Place | New York |
| Access Restriction | One Nation One Subscription (ONOS) |
| Subject Keyword | Benzylisoquinoline alkaloid metabolism Green fluorescent protein Metabolite trafficking Sanguinarine biosynthesis Subcellular localization Cell Biology Plant Sciences Plant Genetics & Genomics Plant Breeding/Biotechnology Developmental Biology |
| Content Type | Text |
| Resource Type | Article |
| Subject | Cell Biology Developmental Biology Plant Science Biotechnology |
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