Open AccessNMR studies of the Escherichia coli trp aporepressor
Author(s) -
HYDE Eva I.,
RAMESH Vasudevan,
ROBERTS Gordon C. K.,
ARROWSMITH Cheryl H.,
TREATCLEMONS Lynda,
KLAIC Branimir,
JARDETZKY Oleg
Publication year - 1989
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1989.tb21083.x
Subject(s) - homonuclear molecule , chemistry , nuclear magnetic resonance spectroscopy , two dimensional nuclear magnetic resonance spectroscopy , dimer , crystallography , nuclear overhauser effect , spectroscopy , chemical shift , stereochemistry , nuclear magnetic resonance , physics , molecule , organic chemistry , quantum mechanics
The resonances in the aromatic region of the 1 H‐NMR spectrum of the Escherichia coli trp aporepressor have been assigned to amino acid type by two‐dimensional correlated spectroscopy (COSY), homonuclear Hartmann‐Hahn (HOHAHA) spectroscopy and nuclear Overhauser enhancement spectroscopy (NOESY) techniques and studies of the pH dependence of the chemical shifts, in combination with selective deuteration of the protein. Complete sequence‐specific assignments of the aromatic resonances have been made by comparing the observed inter‐residue NOEs with those expected on the basis of the crystal structure of the protein [Zhang, R.‐G., Joachimiak, A., Lawson, C. L., Shevitz, R. W., Otwinowski, Z. & Sigler, P. B. (1987) Nature 327 , 591–597]. The latter experiments have also permitted the sequence‐specific assignment of some of the high‐field methyl resonances. The complete assignment of the aromatic region of the spectrum, in particular of resonances from residues at the dimer interface, opens the way to detailed studies of the conformational effects of corepressor and operator binding.