Abstract
Using the perturbed angular correlations (PAC) technique, the formation of hafnium-ovotransferrin complexes has been studied. Two binding configurations at each of the two specific binding-sites of the protein have been observed. They are characterized by well-defined electric quadrupole frequencies. Information about the dynamics of the protein was derived from temperature dependent measurements of the relaxation constant. The well-resolved spectra taken with fast BaF2-detectors allow a precise determination of the relaxation behaviour of the protein. The results are compared with the predictions from a hydrodynamic model for the reorientation of macromolecules. Thus the hydrodynamic volume of ovotransferrin and its N-terminal half-molecule were determined. The ovotransferrin volume is in agreement with a value derived for human serum transferrin from small angle neutron scattering. From experiments with immobilized protein material there is evidence for internal protein dynamics which is probed by the Hf-ion bound to the specific metal-sites.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- PAC:
-
perturbed angular correlations technique
- TF:
-
serum transferrin
- LF:
-
lactoferrin
- OTF:
-
ovotransferrin
- OTF/2N:
-
N-terminal half-molecule of ovotransferrin
- NQR:
-
nuclear quadrupole resonance
- EFG:
-
electric field gradient
- NQI:
-
nuclear quadrupole interaction
- NTA:
-
nitrilotriacetate
- MES:
-
2-(N-morpholino)ethanesulphonic acid
- HEPPS:
-
N-2-hydroxyethyl-piperazine-N′-3-propanesulphonic acid
- TRIS:
-
tris(hydroxymethyl)aminomethane
References
Abola J, Wood MK, Chweh A, Abraham D, Pulsinelli PD (1982) Structure of hen ovotransferrin at 5 Å resolution. In: Saltman P, Hegenauer J (eds) The biochemistry and physiology of iron. Elsevier Biomedical, New York, pp 27–32
Aisen P, Harris DC (1989) Physical biochemistry of the transferrins. In: Loehr TM, Gray HB, Lever ABP (eds) Iron carriers and iron proteins. VCH, Weinheim, pp 239–371
Anderson BF, Baker HM, Norris GE, Rice DE, Baker EN (1989) Structure of human lactoferrin: crystallographic structure analysis and refinement at 2.8 Å resolution. J Mol Biol 209:711–734
Anderson BF, Baker HM, Norris GE, Rumball SV, Baker EN (1990) Apolactoferrin structure demonstrates ligand-induced conformational changes in transferrins. Nature 344:784–787
Appel H, Duffield J, Taylor DM, Then GM, Thies W-G (1987) 181Hf-labelled rat serum transferrin: Influence of temperature on the metal-binding configurations. Hyp Int 35:953–956
Appel H, Neu-Müller M, Brown-Mason A, Schwab F, Taylor DM, Thies W-G (1990) Metal binding in ovotransferrin: should the two observed binding configurations be attributed to one molecular lobe each? Hyp Int 61:1223–1226
Bailey S, Evans RW, Garrat RC, Gorinsky B, Hasnain S, Horsburgh C, Johti H, Lindley PF, Mydin A, Sarra R, Watson JL (1988) Molecular structure of serum transferrin at 3.3 Å resolution. Biochemistry 27:5804–5812
Barfuß H, Böhnlein G, Hohenstein H, Kreische W, Niedrig H, Appel H, Heidinger R, Raudies J, Then GM, Thies W-G (1982) Temperature dependence of the electric field gradient in CdSe and HfO2O2. Z Phys B 47:99–102
Baudry A, Boyer P, Vuillet P (1976) PAC study of molecular reorientation in liquids. Hyp Int 2:279–281
Baudry A, Boyer P, Fabris JD, Vuillet P (1981) PAC study of molecular rotational motions in dilute solutions. Hyp Int 10:1057–1062
Baudry A, Boyer P, Vuillet P (1983) High-resolution TDPAC experiments with fast BaF2 scintillators. Hyp Int 13:263–269
Bauer R (1985) Perturbed angular correlation spectroscopy and its application to metal sites in proteins: possibilities and limitations. Q Rev Biophys 18:1–64
Boyer P, Baudry A (1984) Perturbed angular correlation of gamma rays. In: Matsuura T (ed) Studies in physical and theoretical chemistry 31: hot atom chemistry. Elsevier, Amsterdam
Brock J (1985) Transferrins. In: Harrison PM (ed) Topics in molecular and structural biology, VCH, Weinheim, 7:183–262
Butz T, Lerf A (1983) Comment on ”Mößauer studies of the 6.2 γ-rays of 181Ta in Ta-dichalcogenides”. Phys Lett 97A:217–220
Debye P (1945) Polar molecules, Chap. 5. Dover, New York
van Eijk HG, van Noort WL (1976) Isolation of rat transferrin using CNBr-activated Sepharose 4B. J Clin Chem Clin Biochem 14:475–478
Frauenfelder H, Petsko GA, Tsernoglou D (1979) Temperature-dependent X-ray diffraction as a probe of protein structural dynamics. Nature 280:558–563
Frauenfelder H, Steffen RM (1965) Angular correlations. In: Siegbahn K (ed) Alpha-, beta- and gamma-ray spectroscopy, vol. 2. North Holland, Amsterdam, pp 997–1198
Gorinsky B, Horsburgh C, Lindley PF, Moss DS, Parker M, Watson JL (1979) Evidence for the bilobal nature of diferric rabbit plasma transferrin. Nature 281:157–158
Heidinger R, Thies W-G, Appel H, Then GM (1987) High resolution 181Hf-TDPAC spectroscopy using fast BaF2 detectors. Hyp Int 35:1007–1010
de Jong G, van Dijk JP, van Eijk HG (1990) The biology of transferrin. Clin Chim Acta 190:1–46
Karplus M (1987) Molecular dynamics simulations of protein. Phys Today 40:68–72
Marsden PJ, Smith FA, Evans RW (1989) Evidence of conformational changes in the non-equivalent binding sites of human serum transferrin. Appl Radiat Isol 40:715–722
Marshall AG (1978) Biophysical chemistry: Principle techniques and applications. Wiley, New York, p 528
Martel P, Kim SM, Powell BM (1980) Physical characteristics of human transferrin from small angle neutron scattering. Biophys J 31:371–380
Metz-Boutigue M-H, Jolles J, Mazurier J, Schoentgen F, Legrand D, Spik G, Montreuil J, Jolles P (1984) Human lactoferrin: amino acid sequence and structural comparison with other transferrins. Eur J Biochem 145:659–676
Oe H, Doi E, Hirose M (1988) Amino-terminal and carboxyl-terminal half-molecules of ovotransferrin: preparation by a novel procedure and their interactions. J Biochem 103:1066–1072
Perrin F (1936) Mouvement brownien d'un éllipsoide (II). J Phys Radium 7:1–11
Shinar J, Davidov D, Shaltiel D (1984) Proton NMR study of diffusion in continuous, nonstoichiometric metal-hydrogen systems. Phys Rev B 30: 6331–6363
Smith FA, Lurie DJ, Brady F, Danpure HJ, Kensett FJ, Osman S, Silvester DJ, Waters SL (1984) PAC study of 111In binding to transferrin, tropolone and acetylacetone in aqueous solutions. Int J Appl Radiat Isot 35:501–506
Tanford C (1961) Physical chemistry of macromolecules. Wiley, New York
Taylor DM, Lehmann M, Planas-Bohne F, Seidel A (1983) The metabolism of radiohafnium in rats and hamsters: a possible analog of plutonium for metabolic studies. Radiat Res 95:339–358
Taylor DM, Thies W-G, Appel H, Neu-Müller M, Schwab F, Weber HW (1988) Spectroscopic studies of hafnium-transferrin-complexes: differences between the two binding-sites. In: Hamer DH, Winge DR (eds) Metal ion homeostasis: molecular biology and chemistry, Vol. 98. New Series. Alan R Liss, New York, pp 179–188
Then GM, Appel H, Raudies JH, Thies W-G, Duffield J, Taylor DM (1983) The binding of hafnium of nitrilotriacetate. Hyp Int 16:889–892
Author information
Authors and Affiliations
Additional information
Correspondence to: F. J. Schwab.
Rights and permissions
About this article
Cite this article
Schwab, F.J., Appel, H., Neu, M. et al. Influence of protein dynamics on the metal-sites of ovotransferrin. Eur Biophys J 21, 147–154 (1992). https://doi.org/10.1007/BF00185429
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00185429