Isolation and profiling of integral membrane proteins from developing Brassica napus [abstract]

Abstract

During seed development the concentration of starch, oil, and protein fluctuates. At the early stages of seed filling in oilseeds starch is the principal component. Oil (triacylglycerol) and protein concentrations do not reach a maximum until the later stages of seed development. This metabolic shift within the seed, from production of starch to production of oil and protein, indicates that seed metabolism is regulated temporally. To better understand these metabolic changes it is useful to examine the cognate changes in protein expression. Integral membrane proteins represent one class of proteins which are important for inter-organellar metabolic flow. Current two-dimensional electrophoresis techniques are unsuitable for the profiling of hydrophobic membrane proteins. To specifically characterize this class of proteins, this project was focused on developing a reproducible protocol for membrane protein isolation that can be used with standard sodium dodecyl sulfate polyacrylamide gel electrophoresis. Three different techniques for the isolation of integral membrane proteins were compared, and the resulting gels will be presented.