Abstract
The paramagnetic effect of a spin-labeled sulfonyl fluoride, 4-(2,2,5,5-tetramethylpyrrolidine-1-oxyl)-p-fluorosulfonylbenzamide (p-V), when bound to the active site serine residue of the proteases, bovine plasma-activated protein C (APC) and des(1–41)-light-chain-activated protein C (GDAPC), on the longitudinal relaxation rate (T1) of Tl+ bound to these same proteins has been examined by 205Tl+-NMR spectroscopy. The substantial shortening by bound p-V of the T1 for Tl+ has been employed to estimate the distances between Tl+ and the unpaired electron on each protein surface. Assuming that a single cation-binding site exists on each enzyme, electron-nuclear distances of 3.4–3.9 Å have been calculated for each protein. This suggests that the removal of 41 amino acid residues and, concomitantly, all γ-carboxyglutamic acid, from the amino-terminal of the light chain of APC, does not significantly affect the protein topography in the region of the molecule probed by this technique.
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Hill, K.A.W., Castellino, F.J. Topographical relationships among the monovalent cation binding sites of bovine plasma-activated protein C and des(1–41)-light-chain-activated bovine plasma protein C and a nitroxide spin label bound to their active site serine residues. J Protein Chem 6, 489–495 (1987). https://doi.org/10.1007/BF00276735
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DOI: https://doi.org/10.1007/BF00276735