Multinuclear magnetic resonance studies of metal ion binding sites of phosphoglucomutase.
Metal binding at the activating site of rabbit muscle phosphoglucomutase has been studied by 31P, 7Li, and 113Cd NMR spectroscopy. A 7Li NMR signal of the binary Li+ complex of the phosphoenzyme was not observed probably because of rapid transverse relaxation of the bound ion due to chemical exchange with free Li+. The phosphoenzyme-Li+-glucose 6-phosphate ternary complex is more stable, kinetically, and yields a well-resolved peak from bound Li+ at -0.24 ppm from LiCl with a line width of 5 Hz and a T1 relaxation time of 0.51 +/- 0.07 s at 78 MHz. When glucose 1-phosphate was bound, instead, the chemical shift of bound 7Li+ was -0.13 ppm; and in the Li+ complex of the dephosphoenzyme and glucose bisphosphate a partially broadened 7Li+ peak appeared at -0.08 ppm. Thus, the bound metal ion has a somewhat different environment in each of these three ternary complexes. The 113Cd NMR signal of the binary Cd2+ complex of the phosphoenzyme appears at 22 ppm relative to Cd(ClO4)2 with a line width of 20 Hz at 44.4 MHz. Binding of substrate and formation of the Cd2+ complex of the dephosphoenzyme and glucose bisphosphate broaden the 113Cd NMR signal to 70 Hz and shift it to 75 ppm. The 53 ppm downfield shift upon the addition of substrate along with 1H NMR data suggests that one oxygen ligand to Cd2+ in the binary complex is replaced by a nitrogen ligand at some intermediate point in the enzymic reaction.(ABSTRACT TRUNCATED AT 250 WORDS)
Rhyu
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Ray
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Markley
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D000818:Animals; D001665:Binding Sites; D002104:Cadmium / Q000378:metabolism*; D002712:Chlorides / Q000378:metabolism*; D005958:Glucosephosphates / Q000378:metabolism; D007700:Kinetics; D008094:Lithium / Q000378:metabolism*; D018021:Lithium Chloride; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D009132:Muscles / Q000201:enzymology; D010733:Phosphoglucomutase / Q000378:metabolism*; D011485:Protein Binding; D011817:Rabbits
D002712:Chlorides; D005958:Glucosephosphates; D002104:Cadmium; D008094:Lithium; D010733:Phosphoglucomutase; D018021:Lithium Chloride