Identification of enzymes involved in maturation of plant signaling peptides at Københavns Universitet

Identification of enzymes involved in maturation of plant signaling peptides

Background:

The highly complex network of cell-to-cell communication is crucial for the regulatory mechanisms involved in plant development and stress responses. Signaling peptides are involved in many aspects of plant regulation, including developmental processes and responses to biotic and abiotic stress factors.

When we understand the physiological role of specific signaling peptides, they can potentially be used as biological growth regulators.

Signaling peptides are derived from prepropeptides, which are precursor peptide sequences of approximately 70–200 amino acids.

Post-translationally modified peptides are modified and processed in the secretory pathway before secretion, typically modifications can be tyrosine sulfation, proline hydroxylation and hydroxyproline arabinosylation

We offer two projects related to PSY-peptide maturation:

1)Identification of subtilase responsible for the final processing of PSY1 peptide

2)Characterisation of prolinehydroxylases and identification of the isoform targeting PSY1 peptide

Methods that can used:

Project 1

  • Prediction of candidates by bioinformatics
  • Expression and purification of candidate enzymes in

E.coli and test on PSY1-GST/GST substrate

  • Activity based enzyme purification with PSY1-GST/GST substrate

Project 2

  • Expression and purification of candidate genes in

Pichia pastoris or Saccharomyces. cerevisae (we have the 13 genes synthesized and Pichia strains stably transformed with 10 of them)

  • Activity test on P-rich peptides (Crude assay, MassSpec based assay)

Keywords:
Protein purification, post translationally modification, PTM, heterologous expression, enzyme characterisation

Supervisors:

You will be associated with a postdoc or PhD student as co-supervisor in addition to your main supervisor, Anja Fuglsang.

Attention: Often you need a pre-approval from your university or study counselor, to ensure that projects or thesis found on AU Job- og Projektbank will be accepted as part of your education. Please contact the right entity in due time to ensure that you're picking the right project.