BSc projects about flippases

The search for a biological partner

Tools for detection of protein-protein interactions

Protein-protein interactions can be extremally important in biological systems, as they control the activation or inactivation of catalytically-relevant proteins or even full metabolic pathways. These interactions can be tested using several different techniques. Initially, the yeast-2-hybrid  system was used for this purpose. In this, a transciption factor is split into an activation domain and a DNA-binding domain, that are fused independently to two soluble proteins of interest. If the problem-proteins interact, the two transcription factor domains can work together to activate transcription of a series of reporter genes. Later on, several modifications of this method were developed to test for interaction between membrane proteins (such as the split-ubiquitin system) or to improve the sensitivity of the detection system (like the split Yellow Fluorescent Protein technique). In very recent years, several other approaches have been described that allow for high throughput screening (antibody arrays, Biomolecular Mass Spectrometry). The objective of this bachelor project is to make a comprehensive review of the available techniques for detecting protein-protein interactions. Optionally, short practical work with some of these systems can be performed. The work will be carried out at the PUMPKIN Centre of Excellence in the Faculty of Life Sciences. For information contact Rosa Lopez ()

 

In and out: understanding coordinated cellular processes in the secretory pathway.

Inhibitors and drugs as tools in cell biology
The plant secretory pathway is a highly complex machinery, in which several protein families work in a coordinated way to generate and fuse vesicles to and from different organelle membranes. This coordination is vital for the cells, as it allows the proteins and substrates required for a concrete process (e.g. a defense response under pathogen attack) to come together at the right place and time (e.g., the plasma membrane during penetration of the pathogen). To understand the dynamics of the secretory pathway, not only in plants, but also in other organisms, it is possible to use substances that interfere with vesicle production in different membrane compartments (such as wortmannin or befeldrin A). This way, a part of the secretory pathway can be studied in an isolated way, e.g. uptake of substances by endocytosis can be disrupted without affecting secretion outside the cells. The aim of this bachelor project is to analyse the effect of different inhibitory substances in the functioning of the secretory pathway, and to compare these effects with those observed in mutants with a defective secretory or endocytic machinery. The work will be carried out at Flippase group the PUMPKIN Centre of Excellence in the Faculty of Life Sciences. In the Flippase group we are interested on understanding the physiological role and mechanism of proteins involved in the process of vesicle formation. For information contact Rosa Lopez )

 

Senescence and the hypersensitive response in plants: are flippases the link?

Several types of programmed cell death occur in plants, among others, senescence (the last stage in leaf development) and the hypersensitive response to plant-pathogen interactions. An overlap between the pathogen-response and senescence programs is beginning to be characterized. In our group we are interested in membrane proteins named lipid flippases: proteins that move lipids from one side of a biological membrane to the other using ATP as a driving force. Recently, together with some collaborators, we discovered a possible relationship between flippases and the two above mentioned forms of program cell death. The objective of this project is to learn more about programe cell death in plants, with a focus on senescence and pathogen response and discuss the possible implication of flippases in these process. Some hands-on work could also be included, so come pay a visit if you find the project interesting!!

The work will be carried out at Flippase group the PUMPKIN Centre of Excellence in the Faculty of Life Sciences. In the Flippase group we are interested on understanding the physiological role and mechanism of proteins involved in the process of vesicle formation. For information contact Rosa Lopez ()