Direct real-time observation of actin filament branching mediated by Arp2/3 complex using total internal reflection fluorescence microscopy
AUTOR(ES)
Amann, Kurt J.
FONTE
The National Academy of Sciences
RESUMO
Existing methods for studying actin filament dynamics have allowed analysis only of bulk samples or individual filaments after treatment with the drug phalloidin, which perturbs filament dynamics. Total internal reflection fluorescence microscopy with rhodamine-labeled actin allowed us to observe polymerization in real time, without phalloidin. Direct measurements of filament growth confirmed the rate constants measured by electron microscopy and established that rhodamine actin is a kinetically inactive tracer for imaging. In the presence of activated Arp2/3 complex, growing actin filaments form branches at random sites along their sides, rather than preferentially from their barbed ends.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=64974Documentos Relacionados
- Importance of free actin filament barbed ends for Arp2/3 complex function in platelets and fibroblasts
- Arp2/3 complex requires hydrolyzable ATP for nucleation of new actin filaments
- Arp2/3 Complex from Acanthamoeba Binds Profilin and Cross-links Actin Filaments
- Scar, a WASp-related protein, activates nucleation of actin filaments by the Arp2/3 complex
- The interaction of Arp2/3 complex with actin: Nucleation, high affinity pointed end capping, and formation of branching networks of filaments
