Proteins vanadyl

Chasteen ND, DeKoch RJ, Rogers BL, Hanna MW. 1973. Use of the vanadyl(IV) ion as a new spectroscopic probe of metal binding to proteins vanadyl insulin. J Am Chem Soc 95 1301-1309. 

Insulin binding to the extracellular side of cell membranes initiates the insulin cascade , a series of phosphorylation/dephosphorylation steps. A postulated mechanism for vanadium is substitution of vanadate for phosphate in the transition state structure of protein tyrosine phosphatases (PTP).267,268 In normal physiological conditions, the attainable oxidation states of vanadium are V111, Viv and Vv. Relevant species in solution are vanadate, (a mixture of HV042-/ H2VOO and vanadyl V02+. Vanadyl is not a strong inhibitor of PTPs, suggesting other potential mechanisms for insulin mimesis for this cation. 

Vanadate is reduced in red cells to vanadyl ion by intracellular glutathione following uptake through a phosphate transport system.1072,1073 Free vanadyl is normally unstable with respect to oxidation, but appears to be stable when complexed with intracellular proteins or smaller molecules.1074 Vanadyl ion is a relatively powerful inhibitor of (Na+, K+)-ATPase. For pure fractions of the enzyme, inhibition was nearly complete at less than 5 pmol dm-3 vanadyl ion.1075 The state of vanadyl ion at pH 7 is somewhat uncertain, but may involve a hydroxylated species. Vanadyl ion also inhibits alkaline phosphatase more effectively than does vanadate.1076... 

Vanadate transport in the erythrocyte was shown to occur via facilitated diffusion in erythrocyte membranes and was inhibited by 4,4 -diisothiocyanostilbene-2,2 -disulfonic acid (DIDS), a specific inhibitor of the band 3 anion transport protein [23], Vanadium is also believed to enter cells as the vanadyl ion, presumably through cationic facilitated diffusion systems. The divalent metal transporter 1 protein (called DMT1, and also known as Nramp2), which carries iron into cells in the gastrointestinal system and out of endosomes in the transferrin cycle [24], has been proposed to also transport the vanadyl cation. In animal systems, specific transport protein systems facilitate the transport of vanadium across membranes into the cell and between cellular compartments, whereas the transport of vanadium through fluids in the organism occurs via binding to proteins that may not be specific to vanadium. 

The active sites of these enzymes can have a nitrogen ligand, usually as histidine (acid phosphatases and some protein phosphatases), a nucleophilic serine residue (alkaline phosphatases), a cysteine residue in which the thiol group can form a covalent species with the phosphate ester (protein phosphatases), or an aspartate-linked phosphate (plasma membrane ion pumps). The inhibitory form of vanadium is usually anionic vanadate V(V), but cationic vanadyl V(IV) has also shown strong inhibition of some types of phosphorylase reactions. Above neutral pH, speciation of vanadyl ions produces anionic V(IV) species capable of inhibition of enzymes in the traditional transition-state analogue manner [5],... 

Shioda, N., Ishigami, T., Han, F., Moriguchi, S., Shibuya, M., Iwabuchi, Y., and Fukunaga, K. (2007). Activation of phosphatidylinositol 3-kinase/protein kinase B pathway by a vanadyl compound mediates its neuroprotective effect in mouse brain ischemia. Neuroscience 148, 221-229. 

Cannon, J. C., Chasteen, N. D. The Distinction between Metal Binding Sites in Vanadyl Transferrin Complexes in Proteins of Iron Storage and Transport in Biochemistry and Medicine, (ed.) Crichton, R. R, Amsterdam, North Holland Press, 1975... 

While the affinity of transferrin see Iron Proteins for Storage Transport their Synthetic Analogs) for vanadyl is 10-fold greater than that of albumin, the latter can bind up to 20 vanadyl ions including a specific interaction with cys-34, the only reduced cysteine residue in the protein. 

Other enzymes such as the cyclic AMP-dependent protein kinase are stimulated by vanadium. Vanadate seems to inhibit most strongly those enzymes that form a phosphoenzyme intermediate. This inhibition may be diminished within cells because vanadate is readily reduced by glutathione and other intracellular reductants. The resulting vanadyl ion is a much weaker inhibitor and also stimulates several metabolic processes. ... 

Vanadium is an element, and as such, is not metabolized. However, in the body, there is an interconversion of two oxidation states of vanadium, the tetravalent form, vanadyl (V+4), and the pentavalent form, vanadate (V+5). Vanadium can reversibly bind to transferrin protein in the blood and then be taken up into erythrocytes. These two factors may affect the biphasic clearance of vanadium that occurs in the blood. Vanadate is considered more toxic than vanadyl, because vanadate is reactive with a number of enzymes and is a potent inhibitor of the Na+K+-ATPase of plasma membranes (Harris et al. 1984 Patterson et al. 1986). There is a slower uptake of vanadyl into erythrocytes compared to the vanadate form. Five minutes after an intravenous administration of radiolabeled vanadate or vandadyl in dogs, 30% of the vanadate dose and 12% of the vanadyl dose is found in erythrocytes (Harris et al. 1984). It is suggested that this difference in uptake is due to the time required for the vanadyl form to be oxidized to vanadate. When V+4 or V+5 is administered intravenously, a balance is reached in which vanadium moves in and out of the cells at a rate that is comparable to the rate of vanadium removal from the blood (Harris et al. 1984). Initially, vanadyl leaves the blood more rapidly than vandate, possibly due to the slower uptake of vanadyl into cells (Harris et al. 1984). Five hours after administration, blood clearance is essentially identical for the two forms. A decrease in glutathione, NADPH, and NADH occurs within an hour after intraperitoneal injection of sodium vanadate in mice (Bruech et al. 1984). It is believed that vanadate requires these cytochrome P-450 components for oxidation to the vanadyl form. A consequence of this action is the diversion of electrons from the monooxygenase system resulting in the inhibition of drug dealkylation (Bruech et al. 1984). 

Vanadium in the plasma can exist in a bound or unbound form (Bruech et al. 1984). Vanadium as vanadyl (Patterson et al. 1986) or vanadate (Harris and Carrano 1984) reversibly binds to human serum transferrin at two metalbinding sites on the protein. With intravenous administration of vanadate or vanadyl, there is a short lag time for vanadate binding to transferrin, but, at 30 hours, the association is identical for the two vanadium forms (Harris et al. 1984). The vanadium-transferrin binding is most likely to occur with the vanadyl form as this complex is more stable (Harris et al. 

In Table 2.5 (Section 2.2.1), the high molecular mass blood constituents albumin and transferrin are listed along with the main low molecular mass constituents. Transferrin in particular is a very strong binder for the vanadyl ion [log A = 14.3(6)] and more or less replaces other ligands. The coordination of vanadyl (and vanadate) to transferrin will be taken up in the more general context of vanadium-protein interaction (Section 5.2). 

The (metabolic) pathways of dietary vanadium, such as vanadate [H2V04], can be expressed as illustrated in Scheme 5.1 after oral uptake, vanadate reaches the gastrointestinal tract, where it is partially reduced and precipitated to vanadyl (VO ) hydroxides, which are excreted with the faeces. Another portion is absorbed and circulated in the blood, where it undergoes redox speciation and complexation by the serum proteins transferrin and albumin. Vanadate and vanadyl are finally incorporated into cells, mainly those of the liver, spleen and kidney. Excretion is achieved via the urine. Part of the vanadium is taken up by bones, where the mean retention time is comparatively long. 

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