predicting and testing enzymatic activity
DESCRIPTION
Predicting and testing enzymatic activity. Charlie Harrison Supervisors: Profs. John Ward & Christine Orengo. Norcoclaurine. Benzylisoquinoline alkaloids ( BIQAs ) group of plant secondary metabolites. I ncludes many interesting compounds Common entry point: norcoclaurine. - PowerPoint PPT PresentationTRANSCRIPT
![Page 1: Predicting and testing enzymatic activity](https://reader035.vdocuments.site/reader035/viewer/2022062218/56816384550346895dd46985/html5/thumbnails/1.jpg)
Predicting and testing enzymatic activity
Charlie HarrisonSupervisors: Profs. John Ward &
Christine Orengo
![Page 2: Predicting and testing enzymatic activity](https://reader035.vdocuments.site/reader035/viewer/2022062218/56816384550346895dd46985/html5/thumbnails/2.jpg)
Norcoclaurine• Benzylisoquinoline alkaloids (BIQAs)– group of plant secondary metabolites
• Includes many interesting compounds• Common entry point: norcoclaurine
(S)-norcoclaurine (R)-norcoclaurine
![Page 3: Predicting and testing enzymatic activity](https://reader035.vdocuments.site/reader035/viewer/2022062218/56816384550346895dd46985/html5/thumbnails/3.jpg)
Norcoclaurine• NorcoClaurine Synthase – NCS• Five known examples from three different
species• But BIQAs exist in many more species…• Can we find more NCSs?
![Page 4: Predicting and testing enzymatic activity](https://reader035.vdocuments.site/reader035/viewer/2022062218/56816384550346895dd46985/html5/thumbnails/4.jpg)
Main Project Aims• Bioinformatics– Identify uncharacterised proteins that are likely to
exhibit NCS activity• Lab Work– Express these proteins in E.coli and test
predictions
![Page 5: Predicting and testing enzymatic activity](https://reader035.vdocuments.site/reader035/viewer/2022062218/56816384550346895dd46985/html5/thumbnails/5.jpg)
Bioinformatics• Available data: sequence and structure
• Use these to predict function
![Page 6: Predicting and testing enzymatic activity](https://reader035.vdocuments.site/reader035/viewer/2022062218/56816384550346895dd46985/html5/thumbnails/6.jpg)
Bioinformatics
Reproduced from Orengo, Jones & Thornton 2003 with permission
![Page 7: Predicting and testing enzymatic activity](https://reader035.vdocuments.site/reader035/viewer/2022062218/56816384550346895dd46985/html5/thumbnails/7.jpg)
Bioinformatics• Structural data is better for identifying
functional homologues...
![Page 8: Predicting and testing enzymatic activity](https://reader035.vdocuments.site/reader035/viewer/2022062218/56816384550346895dd46985/html5/thumbnails/8.jpg)
Bioinformatics• Structural data is better for identifying
functional homologues...but is only available for ~1% of sequences
• Use predictions of structural regions – Gene3D• Starting point: Gene3D family of likely
structural relatives• Sequence clustering
![Page 9: Predicting and testing enzymatic activity](https://reader035.vdocuments.site/reader035/viewer/2022062218/56816384550346895dd46985/html5/thumbnails/9.jpg)
Bioinformatics
![Page 10: Predicting and testing enzymatic activity](https://reader035.vdocuments.site/reader035/viewer/2022062218/56816384550346895dd46985/html5/thumbnails/10.jpg)
Lab Work• Design and order the synthetic genes
Wait 3 weeks…
![Page 11: Predicting and testing enzymatic activity](https://reader035.vdocuments.site/reader035/viewer/2022062218/56816384550346895dd46985/html5/thumbnails/11.jpg)
Lab Work
![Page 12: Predicting and testing enzymatic activity](https://reader035.vdocuments.site/reader035/viewer/2022062218/56816384550346895dd46985/html5/thumbnails/12.jpg)
Results
✓ ✓
✗
![Page 13: Predicting and testing enzymatic activity](https://reader035.vdocuments.site/reader035/viewer/2022062218/56816384550346895dd46985/html5/thumbnails/13.jpg)
Results
![Page 14: Predicting and testing enzymatic activity](https://reader035.vdocuments.site/reader035/viewer/2022062218/56816384550346895dd46985/html5/thumbnails/14.jpg)
Thanks