Fragmin: a calcium ion sensitive regulatory factor on the formation of actin filaments

Abstract

Physarum actinin previously isolated [Hatano, S., & Owaribe, K. (1976) in Cell Motility (Goldman, R., Pollard, T., & Rosenbaum, J., Eds.) Vol. 3, Book B, p 499, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY] was found to be a 1:1 complex of actin and fragmin which is a regulatory factor in the formation of actin filaments. Since fragmin did not contain a cysteine residue, it was purified from the complex by the selective cleavage of actin with 2-nitro-5-thiocyanobenzoic acid, followed by column chromatography. Fragmin had nearly the same molecular weight as actin, but had a quite different amino acid composition. When added to G-actin before polymerization, fragmin accelerated the initial viscosity increase of actin solutions induced by salts, but kept the final viscosity much lower than normal F-actin. When added to F-actin after polymerization, fragmin drastically reduced the viscosity of actin solutions. In both cases, the final products of reaction of fragmin with actin were short F-actin filaments. The number average length of the filaments decreased with the increasing molar ratio of fragmin to actin, and the length distribution was always exponential. Fragmin required for its activity a concentration of free Ca2+ higher than 10(-6) M. When the concentration of free Ca2+ was lower than 10(-7) M, fragmin affected neither actin polymerization nor F-actin. The regulation by Ca2+ was reversible.

A Ca2+‐sensitive actin regulatory protein from smooth muscle

“…Over the past 5 years, Ca'+-dependent severing proteins have been identified in a number of non-muscle cells and tissues, including macrophage [2], platelets [3], blood plasma [4], microvillus [5], thyroid [6], Physarum [7,8], and Dictyostefium [9]; however, Ca2+-dependent severing and capping proteins have not been isolated from skeletal muscle.…”

A gelsolin‐related actin‐severing protein with fully reversible actin‐binding properties from the tail muscle of crayfish, Astacus leptodactylus

“…A distinct class of actin-binding proteins, initially termed actin modulators [ 5 ] , comprises proteins of very different evolutionary origin like gelsolin in vertebrates [6-91, severin [lo] and fragmin [5,11, 121 in slime moulds or earthworm actin modulator [13] in annelids. Nevertheless, all these proteins are remarkably similar both in structure and in function [ 13 -181.…”

Reconstitution of Ca2+‐sensitive gelation of actin filaments with filamin, caldesmon and calmodulin

“…Thus, gelsolin (M r 91 000) from macrophages [31 ], viUin (M r 95 000) from intestinal epithelial cell microvilli [32], and fragmin (M r 43 000) from Physarum polycephalum [33] have been isolated. However, their mode of action on the actin gelation is distinctly different from that of caldesmon-calmodulin system: (1) In the presence of Ca 2+, these proteins inhibit f'flamin-induced F actin gelation by reducing the average filament length and increasing the actin f'dament number [34][35][36].…”