Tau induces cooperative Taxol binding to microtubules.
Taxol and tau are two ligands that stabilize the microtubule (MT) lattice. Taxol is an anti-mitotic drug that binds beta tubulin in the MT interior. Tau is a MT-associated protein that binds both alpha and beta tubulin on the MT exterior. Both Taxol and tau reduce MT dynamics and promote tubulin polymerization. Tau alone also acts to bundle, stiffen, and space MTs. A structural study recently suggested that Taxol and tau may interact by binding to the same site. Using fluorescence recovery after photobleaching, we find that tau induces Taxol to bind MTs cooperatively depending on the tau concentration. We develop a model that correctly fits the data in the absence of tau, yields the equilibrium dissociation constant of approximately 2 microM, and determines the escape rate of Taxol through one pore to be 1.7 x 10(3) (M x s)(-1). Extension of the model yields a measure of Taxol cooperativity with a Hill coefficient of at least 15 when tau is present at a 1:1 molar ratio with tubulin.
Ross
Jennifer L
JL
Department of Physics, University of California, Santa Barbara, CA 93106, USA.
Santangelo
Christian D
CD
Makrides
Victoria
V
Fygenson
D Kuchnir
DK
D000818:Animals; D000972:Antineoplastic Agents, Phytogenic / Q000737:chemistry / Q000493:pharmacokinetics*; D002417:Cattle; D003627:Data Interpretation, Statistical; D036681:Fluorescence Recovery After Photobleaching; D005456:Fluorescent Dyes; D006801:Humans; D007700:Kinetics; D008870:Microtubules / Q000737:chemistry / Q000378:metabolism*; D017239:Paclitaxel / Q000737:chemistry / Q000493:pharmacokinetics*; D013194:Staining and Labeling; D016875:tau Proteins / Q000737:chemistry / Q000378:metabolism*
D000972:Antineoplastic Agents, Phytogenic; D005456:Fluorescent Dyes; D016875:tau Proteins; D017239:Paclitaxel