Aldolase-localization in cultured cells: cell-type and substrate-specific regulation of cytoskeletal associations.
The role of aldolase as a true F- and G-actin binding protein, including modulating actin polymerization, initiating bundling, and giving rise to supramolecular structures that emanate from actin fibrils, has been established using indirect immunofluorescence, permeabilization of XTH-2 cells and keratocytes, and microinjection of fluorescence-labeled aldolase. In addition, binding to intermediate filaments, vimentin, and cytokeratins has been demonstrated. In permeabilized cells in the presence of fructose-1,6-bisphosphate (20-2000 microM) aldolase shifts from association with actin fibres to intermediate filaments. Plenty of free binding sites on microtubules have been revealed by addition of fluorochromed aldolase derived from rabbit skeletal muscle. However, endogenous aldolase was never found associated with microtubules. Differences in actin polymerization in the presence of aldolase as revealed by pyrene-labeled actin fluorimetry and viscosimetry were explained by electron microscopy showing the formation of rod-like structures (10 nm wide, 20-60 nm in length) by association of aldolase with G-actin, which prevents further polymerization. Upon the addition of fructose-1,6-bisphosphate, G-actin-aldolase mixture polymerizes to a higher viscosity and forms stiffer filaments than pure actin of the same concentration.
Schindler
R
R
Kinematic Cell Research Group, Biocenter, Goethe University of Frankfurt/Main, Frankfurt, Germany.
Weichselsdorfer
E
E
Wagner
O
O
Bereiter-Hahn
J
J
D000199:Actins / Q000378:metabolism; D000818:Animals; D002478:Cells, Cultured; D003599:Cytoskeleton / Q000201:enzymology*; D004727:Endothelium / Q000378:metabolism; D005634:Fructose-Bisphosphate Aldolase / Q000032:analysis / Q000378:metabolism*; D015603:Keratinocytes / Q000378:metabolism; D007700:Kinetics; D022502:Stress Fibers / Q000378:metabolism; D013379:Substrate Specificity; D014982:Xenopus laevis
D000199:Actins; D005634:Fructose-Bisphosphate Aldolase