Assignment of the 1H-NMR spectrum of a lac repressor headpiece-operator complex in H2O and identification of NOEs. Consequences for protein-DNA interaction.
A complex between the headpiece amino-terminal residues 1-56 of lac repressor (HP56) and an 11-bp lac operator fragment was studied by 1H NMR. The sequence specific assignment of the exchangeable and non-exchangeable protons has been accomplished. Several protons have favourable chemical shifts in the complex, therefore new intraprotein NOEs could be found that had not been unambigously identified in the free protein. By comparison, most of these intraprotein NOEs are also present in the spectra of the free headpiece but some are different. Furthermore, several new proteins DNA NOEs could be identified. The NOE between the side-chain amide protons of Gln18 and C5H of C7 confirms the specific contact between these residues which was proposed from genetic experiments [Ebright, R. M. (1985) J. Biomol. Struct. & Dyn. 3, 281-297]. The implications of the new data for the interaction between the lac repressor headpiece and its operator are discussed.
Lamerichs
R M
RM
Department of Chemistry, University of Utrecht, The Netherlands.
Boelens
R
R
Van der Marel
G A
GA
Van Boom
J H
JH
Kaptein
R
R
D004268:DNA-Binding Proteins / Q000737:chemistry*; D007763:Lac Operon; D009682:Magnetic Resonance Spectroscopy; D012097:Repressor Proteins; D014867:Water
D004268:DNA-Binding Proteins; D012097:Repressor Proteins; D014867:Water