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Studies on minispectrins
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The / -minispectrin is a shortened soluble version §of spectrin constructed to study spectrin §interactions in vitro. In this study we constructed §a modified / ÷N-minispectrin, lacking the 37 amino §acid N-terminal extension of the -minispectrin §chain. The original and modified version of the §minispectrin was produced in order to perform §structural and functional studies. Upon production §of the / - and / ÷N-minispectrin proteins, two §versions of each protein were obtained, a dimer, §( / )/ ( / ÷N), and a tetramer, ( / )2/( / ÷N)2.§§The effect of calcium on the binding of spectrin to §actin has for long been a theme of dispute. §Therefore the influence of calcium on the §interaction between actin and the minispectrins were §investigated using two different methods, an §ultracentrifuge actin pull down assay and Biacore®X. §The results from both methods showed that calcium §had no direct effect on the binding of the §minispectrins to the actin.
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