If we plan an experiment to study the beta-tubulin of an organism w.r.t. its capacity to interact with tubulin binding drugs like Taxol, most studies have used the tubulin purified from some kind of biological source and not the heterologously expressed recombinant beta-tubulin of the target organism. Is it related to the difficulties in expressing the beta-tubulin a properly folded form, its expression or some other reason?
Dear Satpal,
Recombinant tubulin is essentially inactive (or even insoluble). Anyway, natural sources (brains of big mammals) contain ~10% tubulin in the cytoplasmic fraction so that you could consider that they are naturally "overexpressed". Collaboration with groups that routinely purify tubulin may be an option.
Dear Antonio
Thanks for the answer! You have provided some useful info. However in case the study i referred to is planned with a microbe, only the target beta-tubulin from such a microbe would required as taking other organisms's b-tub would not yield the insights being sought...thats what i am interested in...any more inputs please?
Thanks
Satpal Singh
'Functional (i.e., assembly-competent) tubulin can be purified
from pig or cow brain tissue through repeated cycles of polymerization,
high-speed sedimentation of MTs, and depolymerization.
Efforts toward isolating recombinant tubulin isotypes are
hampered by the fact that tubulin biosynthesis is quite complex,
involving chaperones that bind to nascent tubulin polypeptides,
cytosolic chaperonins (CCTs) that facilitate folding of intermediate
tubulin monomers, and tubulin-specific folding cofactors that
allow formation of native heterodimers'
This might answer your question, The reference paper can be found below.
Article Isolation of Functional Tubulin Dimers and of Tubulin-Associ...
Thanks Lakshami for your valuable comments and the paper.
there could still be some way out to study the tubulin from a microbe, i guess!
best regards
Satpal Singh
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