On methods for probing structure and function of membrane proteins

Sammanfattning: The determination of structure–function relationships in membrane proteins plays a key role for the understanding of molecular mechanisms in biological systems, and may provide information on how these systems can be modulated by pharmaceuticals. The aim of the work described herein was to develop methods for studying structure and function of membrane proteins, ideally in combination. The ion channel TRPV1 expressed in Chinese hamster ovary (CHO) cells was used as a model system for evaluation of the methods developed, and further investigations of structure–function relationships in TRPV1 were conducted with these methods. First, a new method for studying ion channel activation was developed based on a microfluidic pipette fabricated in poly(dimethylsiloxane). This device is able to generate a confined volume zone in front of the pipette, and can provide chemical dilution series for the measurement of concentration-response curves in pharmacological studies of cells. The pipette supports delivery of multiple solutions, and rapid switching (