[The comparative analysis of subcellular fractionation methods for revealing of alpha-actinin 1 and alpha-actinin 4 in A431 cells].
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.
A V
AV
Bol'shakova
O A
OA
Petukhova
G P
GP
Pinaev
K E
KE
Magnusson
D000185:Actinin / Q000302:isolation & purification* / Q000378:metabolism; D002021:Buffers; D002458:Cell Fractionation / Q000379:methods*; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism; D002467:Cell Nucleus / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D005615:Freezing; D006801:Humans; D011933:Reagent Kits, Diagnostic; D013347:Subcellular Fractions / Q000737:chemistry
C526905:ACTN1 protein, human; C111453:ACTN4 protein, human; D002021:Buffers; D011933:Reagent Kits, Diagnostic; D000185:Actinin