Studies on minispectrins di Elin Soerhus edito da VDM Verlag
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Studies on minispectrins

Expression in E. coli, purification and in vitro interaction studies

Editore:

VDM Verlag

EAN:

9783639157352

ISBN:

3639157354

Pagine:
84
Formato:
Paperback
Lingua:
Tedesco
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Descrizione Studies on minispectrins

The a/ß-minispectrin is a shortened soluble version of spectrin constructed to study spectrin interactions in vitro. In this study we constructed a modified a/ß÷N-minispectrin, lacking the 37 amino acid N-terminal extension of the ß-minispectrin chain. The original and modified version of the minispectrin was produced in order to perform structural and functional studies. Upon production of the a/ß- and a/ß÷N-minispectrin proteins, two versions of each protein were obtained, a dimer, (a/ß)/ (a/ß÷N), and a tetramer, (a/ß)2/(a/ß÷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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