The comparative analysis of subcellular fractionation methods for revealing of alpha-actinin 1 and alpha-actinin 4 in A431 cells

• Published in: T͡S︡itologii͡a Vol. 51; no. 2; p. 122
• Main Authors: Bol'shakova, A V, Petukhova, O A, Pinaev, G P, Magnusson, K E
• Format: Journal Article
• Language: Russian
• Published: Russia (Federation) 2009
• Subjects: Actinin - isolation & purification; Actinin - metabolism; Buffers; Cell Fractionation - methods; Cell Line, Tumor; Cell Membrane - metabolism; Cell Nucleus - metabolism; Cytoplasm - metabolism; Freezing; Humans; Reagent Kits, Diagnostic; Subcellular Fractions - chemistry
• ISSN: 0041-3771

Alpha-actinin 1 and alpha-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 alpha-actinin 1 and alpha-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 alpha-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 alpha-actinin 4 can be found in all cytoplasmic and nuclear subfractions, while alpha-actinin 1 is characterized by cytoplasmic and membrane localization with specificity of its distribution tightly to the nuclear membrane.