Dear all,
I am quite new in cytoskeleton area. Recently I am finding a gene coding F-actin. But then I found there is not a gene called F-actin but Acta1. After searching some information, I think maybe F-actin is just one formed of acta1. And G-actin is another formed of acta1? So actually there is not a gene named F-actin and we can not do a western bot to quantify the F-actin right (only fluorescence intensity available)?
And I am also planning to detect the polarization level of cytoskeleton. Does anyone have any ideas? thank you so much.
Best regards
Hi there,
First question (from uniprot acta1 mouse https://www.uniprot.org/uniprot/P68134)
"Polymerization of globular actin (G-actin) leads to a structural filament (F-actin) in the form of a two-stranded helix. Each actin can bind to 4 others"
So... There is not a gene called F-Actin or G-Actin. Actin is a family of proteins (there are several actins encoded by different genes)
https://www.uniprot.org/uniprot/?query=actin+AND+homo+sapiens&sort=score).
Also from uniprot:
"In vertebrates 3 main groups of actin isoforms, alpha, beta and gamma have been identified. The alpha actins are found in muscle tissues and are a major constituent of the contractile apparatus. The beta and gamma actins coexist in most cell types as components of the cytoskeleton and as mediators of internal cell motility."
The monomeric form of actins is called G- and the filamentous form is called F-.
Second question:
Yes, you can quantify the amounts of F-Actin and G-Actin by western blot, but you need to do fractions. Check these links
Article The Position of the GFP Tag on Actin Affects the Filament Fo...
https://www.tebu-bio.com/blog/2015/08/18/focus-on-actin-g-f-actin-ratio-measured-in-cells/
Cheers,
J
Acta1 codify for alpha-actin. There are 6 different isoforms encoded by different genes. In cells, you can find actin in two states: monomeric or polymerized. Monomeric actin known also as globular or G-actin, can polymerized and is called filamentous or F-actin. Polymerization can be trigger by different signals. Cells keep a pool of G- actin and F-actin with turnover between both. You can do IF staining to differentiate G-and F-actin or by fractions in WB. Thomas D. Pollard published nice papers about actin dynamics.
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