Characterization of protein dynamics in asymmetric cell division by scanning fluorescence correlation spectroscopy

Research output: Contribution to journalResearch articleContributedpeer-review

Contributors

  • Zdeněk Petrášek - , TUD Dresden University of Technology (Author)
  • Carsten Hoege - (Author)
  • Alireza Mashaghi - (Author)
  • Thomas Ohrt - , TUD Dresden University of Technology (Author)
  • Anthony A. Hyman - (Author)
  • Petra Schwille - , Chair of Biophysics (Author)

Abstract

The development and differentiation of complex organisms from the single fertilized egg is regulated by a variety of processes that all rely on the distribution and interaction of proteins. Despite the tight regulation of these processes with respect to temporal and spatial protein localization, exact quantification of the underlying parameters, such as concentrations and distribution coefficients, has so far been problematic. Recent experiments suggest that fluorescence correlation spectroscopy on a single molecule level in living cells has great promise in revealing these parameters with high precision. The optically challenging situation in multicellular systems such as embryos can be ameliorated by two-photon excitation, where scattering background and cumulative photobleaching is limited. A more severe problem is posed by the large range of molecular mobilities observed at the same time, as standard FCS relies strongly on the presence of mobility-induced fluctuations. In this study, we overcame the limitations of standard FCS. We analyzed in vivo polarity protein PAR-2 from eggs of Caenorhabditis elegans by beam-scanning FCS in the cytosol and on the cortex of C. elegans before asymmetric cell division. The surprising result is that the distribution of PAR-2 is largely uncoupled from the movement of cytoskeletal components of the cortex. These results call for a more systematic future investigation of the different cortical elements, and show that the FCS technique can contribute to answering these questions, by providing a complementary approach that can reveal insights not obtainable by other techniques.

Details

Original languageEnglish
Pages (from-to)5476-5486
Number of pages11
JournalBiophysical journal
Volume95
Issue number11
Publication statusPublished - 1 Dec 2008
Peer-reviewedYes

External IDs

PubMed 18805921

Keywords

ASJC Scopus subject areas