Synthesis of a Cytokinin Linked by a Spacer to Dexamethasone and Biotin: Conjugates to Detect Cytokinin-Binding Proteins
Yeast cells expressing cDNA libraries have provided two new approaches to facilitate further identification of cytokinin-binding proteins and receptors. These are the yeast three hybrid (Y3H) system and fluorescence activated cell sorting (FACS). The Y3H system requires a synthetic hybrid ligand com...
| Main Authors: | , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2016-04-01
|
| Series: | Molecules |
| Subjects: | |
| Online Access: | http://www.mdpi.com/1420-3049/21/5/576 |
| _version_ | 1818585711029256192 |
|---|---|
| author | You Wang David S. Letham Peter C. L. John Ren Zhang |
| author_facet | You Wang David S. Letham Peter C. L. John Ren Zhang |
| author_sort | You Wang |
| collection | DOAJ |
| description | Yeast cells expressing cDNA libraries have provided two new approaches to facilitate further identification of cytokinin-binding proteins and receptors. These are the yeast three hybrid (Y3H) system and fluorescence activated cell sorting (FACS). The Y3H system requires a synthetic hybrid ligand comprising an “anchor” moiety (e.g., dexamethasone) linked to a cytokinin via a spacer. In the yeast nucleus, this ligand by binding connects two fusion proteins leading to a reporter gene activation and detection and characterisation of cytokinin binding proteins. Herein is reported the first synthesis of dexamethasone-cytokinin ligands with a spacer linkage. This was attached to the purine ring of 6-benzylaminopurine (BAP) at positions 2, 8 or 9. To achieve this, dexamethasone was modified by periodate oxidation yielding a carboxylic group used for conjugation to the spacer by amide formation. Biotinyl derivatives of cytokinins for FACS included those synthesised by reaction of an activated ester of biotin with 8-(10-amino-decylamino) derivatives of BAP and BAP 9-riboside. Properties of the conjugates and some biological situations where they could be applicable are discussed briefly. |
| first_indexed | 2024-12-16T08:41:24Z |
| format | Article |
| id | doaj.art-9f8ccc9e8a3a4548976618b8add4a0d3 |
| institution | Directory Open Access Journal |
| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-12-16T08:41:24Z |
| publishDate | 2016-04-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj.art-9f8ccc9e8a3a4548976618b8add4a0d32022-12-21T22:37:40ZengMDPI AGMolecules1420-30492016-04-0121557610.3390/molecules21050576molecules21050576Synthesis of a Cytokinin Linked by a Spacer to Dexamethasone and Biotin: Conjugates to Detect Cytokinin-Binding ProteinsYou Wang0David S. Letham1Peter C. L. John2Ren Zhang3School of Biological Sciences, University of Wollongong, Wollongong, NSW 2522, AustraliaResearch School of Biology, Australian National University, Canberra, ACT 0200, AustraliaResearch School of Biology, Australian National University, Canberra, ACT 0200, AustraliaSchool of Biological Sciences, University of Wollongong, Wollongong, NSW 2522, AustraliaYeast cells expressing cDNA libraries have provided two new approaches to facilitate further identification of cytokinin-binding proteins and receptors. These are the yeast three hybrid (Y3H) system and fluorescence activated cell sorting (FACS). The Y3H system requires a synthetic hybrid ligand comprising an “anchor” moiety (e.g., dexamethasone) linked to a cytokinin via a spacer. In the yeast nucleus, this ligand by binding connects two fusion proteins leading to a reporter gene activation and detection and characterisation of cytokinin binding proteins. Herein is reported the first synthesis of dexamethasone-cytokinin ligands with a spacer linkage. This was attached to the purine ring of 6-benzylaminopurine (BAP) at positions 2, 8 or 9. To achieve this, dexamethasone was modified by periodate oxidation yielding a carboxylic group used for conjugation to the spacer by amide formation. Biotinyl derivatives of cytokinins for FACS included those synthesised by reaction of an activated ester of biotin with 8-(10-amino-decylamino) derivatives of BAP and BAP 9-riboside. Properties of the conjugates and some biological situations where they could be applicable are discussed briefly.http://www.mdpi.com/1420-3049/21/5/5766-benzylaminopurine (BAP)BAP-dexamethasone conjugatesBAP-biotin conjugatesyeast three hybridcytokinin-binding proteins |
| spellingShingle | You Wang David S. Letham Peter C. L. John Ren Zhang Synthesis of a Cytokinin Linked by a Spacer to Dexamethasone and Biotin: Conjugates to Detect Cytokinin-Binding Proteins Molecules 6-benzylaminopurine (BAP) BAP-dexamethasone conjugates BAP-biotin conjugates yeast three hybrid cytokinin-binding proteins |
| title | Synthesis of a Cytokinin Linked by a Spacer to Dexamethasone and Biotin: Conjugates to Detect Cytokinin-Binding Proteins |
| title_full | Synthesis of a Cytokinin Linked by a Spacer to Dexamethasone and Biotin: Conjugates to Detect Cytokinin-Binding Proteins |
| title_fullStr | Synthesis of a Cytokinin Linked by a Spacer to Dexamethasone and Biotin: Conjugates to Detect Cytokinin-Binding Proteins |
| title_full_unstemmed | Synthesis of a Cytokinin Linked by a Spacer to Dexamethasone and Biotin: Conjugates to Detect Cytokinin-Binding Proteins |
| title_short | Synthesis of a Cytokinin Linked by a Spacer to Dexamethasone and Biotin: Conjugates to Detect Cytokinin-Binding Proteins |
| title_sort | synthesis of a cytokinin linked by a spacer to dexamethasone and biotin conjugates to detect cytokinin binding proteins |
| topic | 6-benzylaminopurine (BAP) BAP-dexamethasone conjugates BAP-biotin conjugates yeast three hybrid cytokinin-binding proteins |
| url | http://www.mdpi.com/1420-3049/21/5/576 |
| work_keys_str_mv | AT youwang synthesisofacytokininlinkedbyaspacertodexamethasoneandbiotinconjugatestodetectcytokininbindingproteins AT davidsletham synthesisofacytokininlinkedbyaspacertodexamethasoneandbiotinconjugatestodetectcytokininbindingproteins AT petercljohn synthesisofacytokininlinkedbyaspacertodexamethasoneandbiotinconjugatestodetectcytokininbindingproteins AT renzhang synthesisofacytokininlinkedbyaspacertodexamethasoneandbiotinconjugatestodetectcytokininbindingproteins |