Salt effects on the physical properties of the transferrins.
Salts are known to have a pronounced effect on the spectroscopic, thermodynamic and kinetic properties of human serum transferrin. The present study was undertaken to examine the effect of NaCl on the related proteins ovotransferrin and lactoferrin. EPR difference spectroscopy was used to probe changes in the metal site of these proteins. Sodium chloride was found to perturb the g' = 4.3 EPR spectra of both ovotransferrin and lactoferrin but in different ways. The spectrum of ovotransferrin is reduced in amplitude with a broad feature appearing at g' = 4.8 whereas there is a loss of resolution of the doublet feature at the peak of the EPR derivative spectrum for lactoferrin. The increase in the amplitude of the ovotransferrin EPR difference spectrum (spectrum without NaCl minus spectrum with NaCl) as a function of NaCl concentration is suggestive of saturation binding. A Hill plot binding isotherm gave n = 1.87 +/- 0.32 and log K = 1.49 +/- 0.03 for ovotransferrin, where n is the number of C1- ions binding to either one or both iron containing lobes of the protein and K is the overall association constant. Preliminary measurements with lactoferrin gave n = 1.95 +/- 0.34 and log K = 1.41 +/- 0.06. These results are similar to those previously reported for serum transferrin and suggest that Cl- binds to all the transferrins with strong pairwise cooperativity. This binding may reflect a functional role for chloride and other physiological anions in the uptake and release of iron by the transferrins.
Chasteen
N D
ND
Department of Chemistry, University of New Hampshire, Durham 03824, USA.
Grady
J K
JK
Woodworth
R C
RC
Mason
A B
AB
D002712:Chlorides / Q000378:metabolism; D003207:Conalbumin / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism; D004578:Electron Spin Resonance Spectroscopy; D007700:Kinetics; D007781:Lactoferrin / Q000737:chemistry / Q000378:metabolism; D009994:Osmolar Concentration; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D012492:Salts; D012965:Sodium Chloride / Q000494:pharmacology; D014168:Transferrin / Q000737:chemistry* / Q000378:metabolism
D002712:Chlorides; D011994:Recombinant Proteins; D012492:Salts; D014168:Transferrin; D003207:Conalbumin; D012965:Sodium Chloride; D007781:Lactoferrin