Isolation of the amino-terminal fragment of lactose repressor necessary for DNA binding

Abstract
The lac repressor [of Escherichia coli] can be dissected by trypsin into a homogeneous tetrameric core (accounting for residues 60-347), carrying inducer binding activity, and the monomeric amino-terminal peptides (headpieces) accounting for residues 1-59 and 1-51, respectively. This restriction of the action of trypsin on lac repressor is obtained in 1 M Tris-HCl (pH 7.5)-30% in glycerol at 25.degree. C, since only the peptide bonds at lysine-59 and to a lesser extent at arginine-51 are cleaved under these conditions. The headpieces can be purified by gel filtration. They have ordered secondary structure as revealed by circular dichroism studies. The monomeric headpieces show the relatively weak binding to nonoperator DNA but not the highly specific and strong binding to operator DNA typical for tetrameric lac repressor.

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