Inhibition of actin polymerization in S. cerevisiae.
We want to study the role of actin in the transport of different cargos in S. cerevisiae. The best known method would be to use Latrunculin A or B in order to inhibit actin polymerization (Ayscough et al JCB 1997). Because we want to use biochemical assays and detection methods for our cargos, we work with a bigger culture volume than we would have to use for microscopy or in vitro studies and therefore we would need a bigger amount of Lat A or B.
Is there an alternative drug to Lat A or Lat B which is similar effective in vivo but less costly?
Hi Harald,
An alternative inhibitor of actin polymerization is Cytochalasin D, which I believe is cheaper than Lat A and is cell permeable.
Good luck!
Maya
Hi Harald,
Cytochalasin D is cheaper but less specific inhibitor. Latrunculins are more specific (molecular target is known: G-actin) and they are acting already in nanomolar range while for Cytochalasin D it will be likely micromolar range or higher. Consider this to calculate final cost. Consider also inhibitor specificity necessary for your experiments.
best,
Jozef
Dear Harald,
as already mentioned by colleagues, cytochalasins are the alternative. In my hands, cytochalasin B is best. In plant cells it works specifically and it keeps for long time its effect. But as Jozef underlined, check for the final cost.
However, another idea may be to intefere not with actin but with myosins. BDM (butanedione monoxime) is very potent and cheap. You might then give also a chance to it, even as an extra control for actin-based motility.
Good luck,
Manolis
Cytochalasin D is more specific compare with cytochalasin B for mammalian cells and not so expensive. Anyway, if you can compare for S. cer. in preliminary experiments with different concentrations, it would be better.
Note that depending on the experiment, cytochalasin and latrunculin may have different effects. Cytochalasins block plus end elongation of F-actin filaments while latrunculin blocks assembly by sequestering G-actin. A formin mediated mechanism might be inhibited by latrunculin but not cytochalasin. Both rely on other mechanisms for depolymerization to occur. Also note that BDM is a relatively non-specific ATPase inhibitor, not a myosin inhibitor.
David you are right about BDM, yet a backup test with ML-7 will prove the specificity (or not). If the result is OK, then BDM can be used at the quantity desired and at low cost.
Cytochalasins may be cheaper but do note wipe out F-actin as much as latrunculins do.
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