Identification of the metal coordinating residues in the DNA binding domain of the glucocorticoid receptor by 113Cd-1H heteronuclear NMR spectroscopy.
Two-dimensional 1H-113Cd HSQC and relay HSQC experiments were performed on the 113Cd substituted DNA binding domain of the rat glucocorticoid receptor. The results of these experiments combined with sequence-specific assignments allowed the identification of all coordinating cysteines. It was found that C495 and not C500 is the fourth coordinating cysteine in the second zinc-finger. A signal at approximately 2 ppm previously assigned to a epsilon-CH3 of a methionine residue coordinating to a third, weakly bound, cadmium ion, was identified as the C443 beta proton ligating to the metal ion in the first zinc-finger. No indications were found for the presence of a previously suggested third metal ion binding site.
Department of Chemistry, University of Utrecht, The Netherlands.
E
E
Kellenbach
B A
BA
Maler
K R
KR
Yamamoto
R
R
Boelens
R
R
Kaptein
D000595:Amino Acid Sequence; D000818:Animals; D004268:DNA-Binding Proteins / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D011487:Protein Conformation; D051381:Rats; D011965:Receptors, Glucocorticoid / Q000737:chemistry* / Q000235:genetics; D016335:Zinc Fingers / Q000235:genetics
D004268:DNA-Binding Proteins; D011965:Receptors, Glucocorticoid