Predicting and testing enzymatic activity

Predictingand testing enzymatic activity Charlie Harrison Supervisors: Profs. John Ward & Christine Orengo

Norcoclaurine • Benzylisoquinoline alkaloids (BIQAs) • group of plant secondary metabolites • Includes many interesting compounds • Common entry point: norcoclaurine (S)-norcoclaurine (R)-norcoclaurine

Norcoclaurine • NorcoClaurineSynthase – NCS • Five known examples from three different species • But BIQAs exist in many more species… • Can we find more NCSs?

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

Bioinformatics • Available data: sequence and structure • Use these to predict function

Bioinformatics Reproduced fromOrengo, Jones & Thornton 2003 with permission

Bioinformatics • Structural data is better for identifying functional homologues...

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

Bioinformatics

Lab Work • Design and order the synthetic genes Wait 3 weeks…

Lab Work

Results ✗ ✓ ✓

Results

Thanks

Predicting and testing enzymatic activity