Heteronuclear NMR studies of cobalt corrinoids. 20. 31P chemical shift anisotropy of aquacobalamin and its complex with a haptocorrin from chicken serum.
Static light scattering measurements have been used to determine the molecular mass (65.3 kDa) and second virial coefficient (3.66 x 10(-4) mol mL g-2) for the complex between chicken serum haptocorrin (HC) and aquacobalamin (H2OCbl). Dynamic light scattering measurements have also been used to determine the translational diffusion coefficient of the H2OCbl-HC complex as a function of concentration. From the diffusion coefficient at infinite dilution (4.71 x 10(-7) cm2 s-1), the hydrodynamic radius (45.5 A) and rotational correlation time (85.4 ns) have been calculated. Using the latter, and measured values of the 31P NMR linewidths of the H2OCbl-HC complex at several field strengths, a detailed analysis of the 31P nuclear relaxation is possible. The chemical shift anisotropy term from the transverse relaxation equation is determined to be 95.7 ppm, and the average phosphorus-proton internuclear distance is 2.05 A. For comparison to protein-free H2OCbl, the chemical shift anisotropy of the phosphorus atom was studied by solid state NMR spectroscopy and the 31P relaxation by solution T1 measurements. These studies give values of 110.3 ppm for the chemical shift anisotropy term and 2.45 A for the average phosphorus-proton internuclear distance. The results are consistent with a significant change in the conformation of the H2OCbl phosphodiester upon binding to haptocorrin which could be due to a shortening of the axial Co-N bond.
Brown
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KL
Department of Chemistry, Clippinger Laboratory, Ohio University, Athens 45701, USA. brownk3@oak.cats.ohiou.edu
Wilson
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Jacobsen
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D000818:Animals; D016880:Anisotropy; D002645:Chickens; D066298:In Vitro Techniques; D008027:Light; D046911:Macromolecular Substances; D009682:Magnetic Resonance Spectroscopy; D008970:Molecular Weight; D010758:Phosphorus / Q000737:chemistry; D011487:Protein Conformation; D012542:Scattering, Radiation; D014155:Transcobalamins / Q000737:chemistry*; D014805:Vitamin B 12 / Q000031:analogs & derivatives* / Q000737:chemistry
D046911:Macromolecular Substances; D014155:Transcobalamins; C043960:aquacobalamin; D010758:Phosphorus; D014805:Vitamin B 12