title: Bis-ANS as a specific inhibitor for microtubule-associated protein induced assembly of tubulin.
abstract: 5,5'-Bis[8-(phenylamino)-1-naphthalenesulfonate] (bis-ANS), the fluorescent probe which binds to tubulin, inhibits its assembly into microtubules [Horowitz et al. (1984) J. Biol. Chem. 259, 14647-14650]. The results described in this paper demonstrate that bis-ANS is quite distinct from other well-known microtubule inhibitors in its specificity of action. The inhibitory potentials of bis-ANS and its three structural analogues ANS, Prodan [6-propionyl-2-(dimethylamino)naphthalene], and NSA (naphthalenesulfonic acid) have been compared. It is found that they can be arranged in the following order according to their polymerization inhibitory potentials: bis-ANS approximately equal to Prodan much greater than ANS greater than NSA. Interestingly, the naphthalene nucleus is sufficient to cause inhibition of polymerization. Detailed experiments were carried out to examine the mode of assembly inhibition by aminonaphthalenes at the molecular level, using bis-ANS as a representative. It was found that there was little or no effect of bis-ANS on the assembly of tubulin when polymerization was induced by assembly promoters like taxol, DMSO, or glutamate, or on the assembly of subtilisin-digested protein (tubulin S), for all of which half-maximal inhibition could not be achieved even at 120 microM bis-ANS. On the contrary, bis-ANS acts as an inhibitor in the case of MAP- (MAP2 and tau) and poly(L-lysine)-induced assembly of tubulin, with half-maximal inhibitory concentrations ranging from 1.5 to 7.6 microM. Our results place bis-ANS as a novel inhibitor, which seems to specifically inhibit C-termini-mediated assembly. Of all assembly inhibitors known so far, none exhibits such selection.(ABSTRACT TRUNCATED AT 250 WORDS)
authors.affiliation: Department of Biochemistry, Bose Institute, Calcutta, India.
authors.forename: M
authors.initials: M
authors.lastname: Mazumdar
authors.forename: P K
authors.initials: PK
authors.lastname: Parrack
authors.forename: K
authors.initials: K
authors.lastname: Mukhopadhyay
authors.forename: B
authors.initials: B
authors.lastname: Bhattacharyya
mesh_terms: D000817:Anilino Naphthalenesulfonates / Q000494:pharmacology*; D000818:Animals; D001923:Brain Chemistry; D004305:Dose-Response Relationship, Drug; D006041:Goats; D066298:In Vitro Techniques; D008869:Microtubule-Associated Proteins / Q000378:metabolism*; D009391:Nephelometry and Turbidimetry; D011485:Protein Binding / Q000187:drug effects; D014404:Tubulin / Q000378:metabolism*
chemical_list: D000817:Anilino Naphthalenesulfonates; D008869:Microtubule-Associated Proteins; D014404:Tubulin