Predicting and testing enzymatic activity

Charlie HarrisonSupervisors: 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

• NorcoClaurine Synthase – 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 from Orengo, 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

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Results

Thanks