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Nucleic Acids Research, 1982, Vol. 10, No. 12 3791-3802
© 1982

CHEMISTRY

Structure of the complex between lac repressor headpiece and operator DNA from measurements of the orientation relaxation and the electric dichroism

Dietmar Pörschke*, Norbert Geisler* and Wolfgang Hillen+

*Max-Planck-Institut für Biophysikalische Chemie D-3400 Göttingen, Am Fassberg, Postfach 968 +Technische Hochschule Darmstadt, Institüt fur Organische Chemie Petersenstr. 22, D-6100 Darmstadt, FRG

Received March 4, 1982. Revised May 10, 1982. Accepted May 18, 1982.

The complex between lac repressor headpiece and short rodlike DNA fragments containing the lac operator sequence is characterised by measurements of the rotation diffusion. Using the method of eleotric dichroism we measure the rotation relaxation and determine changes in the length of the DNA upon ligand binding with high accuracy. According to these measurements any change in the length of the operator DNA upon binding of the first two headpiece moleoules remains below 1A; the electrio dichroism also remains virtually unchanged. At high degrees of (unapecific) binding we observe an increase in the rotation relaxation time, which is attributed to an Increase of the apparent mean radius of the complex. As a control of our procedure for the determination of length changes we use the Intercalation of ethidium bromide and arrive at an increase of the DNA length per bound ethidium of 3.2Å (at 3.4Å rise per base pair). The results obtained for the headpiece operator complex are not consistent with models assuming large ohanges of the DNA structure or intercalation of tyrosine residues.


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