Dresden 2009 – wissenschaftliches Programm
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BP: Fachverband Biologische Physik
BP 22: Transport Processes and Cellular Trafficking
BP 22.3: Vortrag
Donnerstag, 26. März 2009, 15:00–15:15, ZEU 260
Fluorescence correlation analysis of protein dynamics in dividing C. elegans embryo — •Zdeněk Petrášek1, Carsten Hoege2, Anthony A. Hyman2, and Petra Schwille1 — 1Biophysics group, Biotechnologisches Zentrum, TU Dresden, Germany — 2Max Planck Institute of Molecular Biology and Genetics, Dresden, Germany
We have combined two-photon fluorescence correlation spectroscopy (FCS), scanning FCS (sFCS) and time-lapse imaging to study the localization and motion of several GFP-labelled proteins involved in the asymmetric first division of C. elegans embryo. The diffusion of all investigated proteins in the cytosol, where they are distributed homogeneously on the scale of optical resolution, was measured with a standard FCS, yielding a distribution of diffusion coefficients. The comparison of the protein size and the obtained diffusion coefficients indicates hindered diffusion or formation of larger complexes.
Two of the investigated proteins, known to play an essential role in the first asymmetric division, PAR-2 and NMY-2, are non-uniformly distributed on the embryo cortex. Their motion was characterized by spatio-temporal correlation measured with sFCS. Scanning FCS reduces the effects of dye photobleaching and improves the statistical accuracy, making it possible to study even slow protein dynamics. The PAR-2 cortical pattern is less concentrated into discrete spots and more dynamic than that of NMY-2, indicating predominantly independent localization of the two proteins on the cortex.