Prof. Andrew Simkin
From
In residence at
Biomolecule and Plant Biotechnology (BBV), University of Tours - FR
Host scientist
Dr Marc Clastre
Biography
Andrew Simkin is a lecturer in the School of Life Sciences at the University of Esses in the United Kingdom. After completing his undergraduate degree at the John Moores University of Liverpool, he earned his Doctor of Philosophy degree from the Université Joseph Fourier, Grenoble, France. With over 20 years of experience in Primary and Secondary metabolism, Dr Simkin has work in both academia and Industry. In Industry, Dr Simkin has worked at Syngenta (Bracknell, UK), Nestle Research and Development (Tours, France) and for the National Institute of Agricultural Botany (NIAB; Kent, UK). He has further experience in several academic environment, working on fruit flavour and production at the University of Florida (Florida, US), on the production of medicinal alkaloids at the Université François-Rabelais (Tours, France), on improving primary metabolism an crop yield at the University of Essex (Colchester, UK). Dr Simkin has a long interest in nutrition and crop production. On becoming an independent Group Leader, he applied his expertise to increasing the yield of fruiting crops and the improvement of the nutritional quality of fruits.
Project
Methods development for proteomic profiling of prenylated proteins
In plants, terpenoids (or isoprenoids) are synthesized by two different pathways: the cytosolic mevalonate (MVA) pathway and the plastidial 2C-methyl-D-erythritol 4-phosphate (MEP) pathway. The particularity of plants to produce isoprenoids via these two pathways, prompts us to address the question of how they might interact and what mechanisms might be involved in coarse control and even fine-tuning of one or both pathways, depending on the structure and functional destination of terpenoid derivatives.
Previous studies performed by our group suggest that prenylated proteins exert specific functions in this problematic. The observations made on monoterpene indole alkaloid biosynthesis regulation in periwinkle (Catharanthus roseus) cell suspensions suggest that one or more prenylated proteins can exert a positive control on the MEP pathway. This view not only proposes an original role of prenylated proteins as a regulator of a biosynthetic pathway, but also raises very important questions as to the relation between MVA and MEP pathways. To shed more light on this question it will be necessary to identify the proteins targeted for prenylation (defined as the prenylome).
The objective of this project is to develop methods for proteomic profiling of periwinkle prenylated proteins. Identification of the prenylome will be investigated by a strategy based on activity-based protein profiling (ABPP). It will be necessary to synthesize the activity-based probes and the reporter tag that will be used to develop the ABPP method. Moreover, experiments using definite targets of prenylation (dansylated peptides and recombinant proteins) will be performed (i) to test if the synthetic activity-based probes are efficient substrates for protein prenylation in vitro (ii) to test the efficiency of the reporter tag for affinity purification of the modified prenylated proteins and (iii) to analyse these proteins by tandem mass spectrometry.
The availability of the specific proteomic probes and the data from the set of experiments will provide essential skills and knowledge for future studies on the identification of the periwinkle prenylome.