Binding of vanadate to human erythrocyte ghosts and subsequent events.
Spectroscopic techniques were used to investigate the interaction between vanadate and human erythrocyte ghosts. Direct evidence from 51 V nuclear magnetic resonance (NMR) studies suggested that the monomeric and polymeric vanadate species may bind to the anion binding sites of band 3 protein of the erythrocyte membrane. The results of 51V NMR studies and the quenching effect of vanadate on the intrinsic fluorescence of the membrane proteins indicated that in the low concentration range of vanadate (< 0.6 mM), monomeric vanadate binds mostly to the anion sites of band 3 protein with the dissociation constant close to 0.23 mM. The experiments of sulfhydryl content titration by the method of Ellman and residue sulfhydryl-labeled fluorescence spectroscopies clearly displayed that vanadate reacts directly with sulfhydryl groups. The appearance of the anisotropic election spin resonance (ESR) signal of vanadyl suggests that a small (c.3%) amount of vanadate was reduced by sulfhydryl groups of membrane proteins. The fluidity and order of intact ghost membrane were reduced by the reaction with vanadate, as shown by the ESR studies employing the protein- and lipid-specific spin labels. It was concluded that although vanadates mainly bind to band 3 protein, a minor part of vanadate may oxidize the residue sulfhydryl groups of membrane proteins, and thus decrease the fluidity of erythrocyte membrane.
Zhang
B
B
Inorganic Chemistry Department, School of Pharmaceutical Sciences, Beijing Medical University, China.
Ruan
L
L
Chen
B
B
Lu
J
J
Wang
K
K
D001457:Anion Exchange Protein 1, Erythrocyte / Q000737:chemistry / Q000378:metabolism; D001665:Binding Sites; D004578:Electron Spin Resonance Spectroscopy; D004910:Erythrocyte Membrane / Q000378:metabolism*; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D008560:Membrane Fluidity; D008565:Membrane Proteins / Q000378:metabolism*; D010084:Oxidation-Reduction; D011108:Polymers; D013050:Spectrometry, Fluorescence; D013438:Sulfhydryl Compounds / Q000737:chemistry / Q000378:metabolism; D014638:Vanadates / Q000737:chemistry / Q000378:metabolism*
D001457:Anion Exchange Protein 1, Erythrocyte; D008565:Membrane Proteins; D011108:Polymers; D013438:Sulfhydryl Compounds; D014638:Vanadates