THE COMPARATIVE ANALYSIS OF SUBCELLULAR FRACTIONATION METHODS FOR REVEALING OF α-ACTININ 1 AND α-ACTININ 4 IN A431
CELLS
A. V. Bolshakova,1 O. A. Petukhova,1 G. P. Pinaev,1 K.-E. Magnusson 2
1 Institute of Cytology RAS, St. Petersburg, and 2 Linkoping University, Linkoping, Sweden;
1 e-mail: asyab@mail.ru
α-Actinin 1 and α-actinin 4 belong to a family of actin-binding proteins with shared structural function and regulation of several processes in a cell.
Based on previous data on different distribution of these proteins in the nucleus and cytoplasm, we have explored in detail the distribution of α-actinin 1 and
α-actinin 4 in subcellular fractions in A431 cells spread on fibronectin. Several methods of subcellular fractionation were used. Complex approach allowed
resuming that revealing of α-actinin isoforms in fractions depended on the composition of lysis buffer and preliminary low-temperature freezing of the cells. We
have drawn a conclusion that α-actinin 4 can be found in all cytoplasmic and nuclear subtractions, while α-actinin 1 is characterized by cytoplasmic and
membrane localization with specificity of its distribution tightly to the nuclear membrane.
Key words: α-actinin 1, α-actinin 4, subcellular fractionation
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