Interaction of microtubule-associated protein with actin filaments. Studies using the fluorescence-photobleaching recovery technique

T. Arakawa, C. Frieden

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8 Scopus citations

Abstract

The interaction of microtubule-associated proteins with actin filaments has been investigated by measuring the diffusion coefficient of either the filament or the microtubule-associated proteins. Experiments were performed using the technique of fluorescence photobleaching recovery with actin labeled with iodoacetamidotetramethyl rhodamine or microtubule-associated proteins labeled with iodoacetamidofluoroescein. Actin filaments composed of pure rhodoamine-labeled actin are not immobilized under a variety of conditions (Tait, J.F., and Frieden, C. (1982c) Biochemistry 21, 6046-6053). We find that addition of microtubule-associated proteins to rhodamine-labeled actin in a ratio as low as 1:1000 can cause immobilization, presumably cross-linking actin into a network of nondiffusible filaments. Immobilization occurs after polymerization is complete, suggesting either a length redistribution of actin filaments, a redistribution of the cross-links between filaments, or the slow addition of actin filaments to other filaments via the microtubule-associated protein. Experiments using fluoresceIn-labeled microtubule-associated proteins show that these proteins are bound to actin filaments as they are formed and that binding depended on actin concentration, indicating that there are a number of binding sites on the actin filaments. However, while the actin filaments become completely immobilized, the microtubule-associated proteins become only partially immobilized suggesting at least two different classes of binding affinities. The large peptide obtained from trypsin-treated fluorescein-labeled microtubule-associated proteins is not able to immobilize actin filaments since it does not bind to the filaments.

Original languageEnglish
Pages (from-to)11730-11734
Number of pages5
JournalJournal of Biological Chemistry
Volume259
Issue number19
StatePublished - Jan 1 1984

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