Francesco Orsini

Atomic Force Microscopy Applied to Biological Systems: Study of Xenopus Laevis Oocyte Plasma Membrane Francesco Orsini

This book describes the application of Atomic Force Microscopy (AFM) to the study of the Xenopus laevis oocyte plasma membrane. In particular, different sample preparation protocols developed and optimized to perform AFM investigation of both external and intracellular sides of the oocyte native plasma membrane are presented and discussed in details. AFM imaging allowed visualization and dimensional characterization of protein complexes observed on both sides of the oocyte plasma membrane. In addition, a new methodological approach based on the purification of oocyte plasma membrane by ultracentrifugation on a discontinuous sucrose gradient allowed to image oocyte membrane in a physiological-like environment as well as to identify a membrane protein, the human aquaporin 4, expressed in the oocytes thus opening interesting perspectives in the AFM investigation of heterologous proteins of relevant biomedical interest. Taken together, these results confirm the potential of AFM as a useful tool for the structural characterization of proteins in native eukaryotic membranes as well as its relevance for describing the organization of protein complexes in native biological membranes.