Biology Reference
In-Depth Information
CHAPTER
on Biological Membranes
19
Robert V. Stahelin
*
,{
Monitoring Peripheral
Protein Oligomerization
*
Department of Biochemistry and Molecular Biology, Indiana University
School of Medicine-South Bend, South Bend, Indiana, USA
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Department of Chemistry and Biochemistry, University of Notre Dame,
Notre Dame, Indiana, USA
CHAPTER OUTLINE
Introduction ............................................................................................................ 360
19.1 Materials and Methods....................................................................................362
19.1.1 Cell Maintenance, Transfection, and Observation .......................... 362
19.1.2 Cellular Imaging and Analysis...................................................... 363
19.1.3 Imaging and Analysis at the Plasma Membrane............................. 363
19.2 Considerations................................................................................................365
Summary and Conclusion ........................................................................................ 366
Acknowledgments ................................................................................................... 369
References ............................................................................................................. 369
Abstract
Peripheral proteins transiently interact with cellular membranes where they regulate
important cellular events such as signal transduction. A number of peripheral pro-
teins harbor lipid-binding modules that not only bind selectively with nanomolar af-
finity to biological membranes but also oligomerize on the membrane surface.
In some cases, specific lipid binding or specific lipid compositions can induce
peripheral protein oligomerization on cellular membranes. These oligomers serve
different roles in biological signaling such as regulating protein-protein interactions,
induction of membrane bending, or facilitating membrane scission. A number of
technologies have been employed to study protein oligomerization with fluctuation
analysis of fluorescently labeled molecules recently developed for use with commer-
cial laser-scanning microscopes. In this chapter, the approach of raster image corre-
lation spectroscopy coupled with number and brightness (N&B) analysis to
investigate protein oligomerization on cellular membranes in live cells is presented.
