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. 1973 Apr;70(4):1007–1011. doi: 10.1073/pnas.70.4.1007

Binding of 13C-Enriched α-Methyl-D-Glucopyranoside to Concanavalin A as Studied by Carbon Magnetic Resonance

C F Brewer *,†,‡,§,, H Sternlicht *, D M Marcus *,†,‡,§,, A P Grollman *,†,‡,§,
PMCID: PMC433413  PMID: 4515601

Abstract

Binding of α-methyl-D-glucopyranoside, uniformly enriched with 14% 13C, to zinc and manganese derivatives of concanavalin A at pH 5.6 was studied by pulsed Fourier transform carbon magnetic resonance techniques. Spin-lattice relaxation (T1) of the [13C]carbons of the sugar was measured in the absence and presence of the two transition metal derivatives of the protein. In the presence of the manganese derivative of concanavalin A, selective relaxation of the sugar carbons was observed. The values for T1 reflect different distances between the carbons of the bound sugar and the manganese ion. Calculation of the distance between the manganese ion and each carbon of the sugar permit the 3-dimensional orientation of the bound sugar to be determined relative to the transition metal site in the protein. The results indicate that α-methyl-D-glucopyranoside binds to the protein in the Cl chair conformation with its 3- and 4- carbons closest to the manganese ion at a mean distance of 10 Å.

Keywords: carbon relaxation, metalloprotein, binding orientation

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Selected References

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