Proteins on the fly

Efficient ER protein folding, secretion, and membrane protein production

PROGRAM

14:00 – 14:40
Mikael Rørdam Andersen, DTU
Protein Secretion Engineering Through Network Approaches
In utilizing living mammalian cells for the production of recombinant proteins, it quickly becomes apparent that the protein secretion pathway is a highly complex combination of cellular organelles and machinery. There are multiple critical points and potential bottlenecks throughout the pathway, going through efficient expression of the gene of interest, correct translocation of the transcript to the endoplasmatic reticulum (ER), peptide synthesis, protein folding and stress responses, protein glycosylation in the Golgi and finally secretion. To understand the interplay of these components, we have adapted and developed multiple systems-­‐‑level analysis methods, to interpret the complex data, characterize individual components, and identify target for engineering in the pathway. Here, I will present some of these methods from our current, coming and previous work

14:40 – 15:20
Neil Bulleid, University of Glasgow
Creating the optimal environment for protein folding in the endoplasmic reticulum
The folding and assembly of proteins within the endoplasmic reticulum (ER) is an essential process for normal healthy tissues. Breakdown in this process leads to several chronic diseases so it is critical that we have a better understanding of the molecular details of how cells fold proteins that enter the secretory pathway. Nearly a third of all proteins coded for by the human genome enter the secretory pathway and undergo disulfide formation during their folding. We know that members of the protein disulfide isomerase family catalyze disulfide formation but we know little about the function of each of the family members or how the unique redox environment of the ER is maintained. In this presentation I will review our current work to characterize the process of correct folding and disulfide formation

Short break

15:30 – 16:10
Jason Schnell, Oxford University
Insights into a membrane integral chaperone: the Sigma‑1 Receptor
The Sigma-1 Receptor (S1R) is a small, multifunctional membrane protein that has chaperone activity and also modulates the activities of ion channels and receptors in a ligand-­‐‑dependent manner. Despite its role in cellular stress responses and as a therapeutic target for pain and addiction few molecular details of S1R have been available. We will report on our progress in applying solution NMR spectroscopy and other biophysical approaches to provide insights into this enigmatic receptor

16:10 – 16:50
Per Amstrup Pedersen, University of Copenhagen
Towards structures of human membrane proteins
Recombinant membrane proteins are notoriously difficult to produce to high membrane densities. Purification is furthermore complicated due to requirements of detergents or amphipols to protect the hydrophobic transmembrane proteins. We have managed to develop a S. cerevisiae based platform with the capacity to deposit eukaryotic membrane proteins to a density of up till 8% of total membrane protein content and identified conditions that allow efficient purification of high quality eukaryotic membrane proteins. In my present talk I will focus on some of our recent results with production and purification of functional human K-channels.