Effects of perturbants on the pink (reduced) active form of uteroferrin. Phosphate-induced anaerobic oxidation.
The binuclear iron cluster of uteroferrin in its reduced and enzymatically active pink form is sensitive to a variety or perturbants. Orthophosphate, in the presence or absence of oxygen, rapidly shifts the absorption maximum of pink uteroferrin from 510 to 545 nm, concurrently abolishing the protein's g'av = 1.74 EPR signal. Apparently, therefore, dioxygen is not required for phosphate-induced oxidation of the pink protein's ferrous iron. Pyrophosphate and arsenate produce changes which differ only in degree from those induced by phosphate, suggesting that all of these structurally similar competitive inhibitors bind to a common site. Molybdate, an inhibitor even more potent than phosphate, quantitatively converts the rhombic EPR signal of pink uteroferrin into an axial signal that remains invariant to subsequent additions of phosphate. Thus, there can be inhibition without oxidation, as further evidenced by the complex EPR spectrum of undiminished intensity produced by sulfate. Fluoride, too, induces an axial component in the EPR signal of pink uteroferrin, but at high concentration abolishes the signal entirely. Vanadate also drives the protein to its oxidized, EPR-silent state, serving as an electron acceptor itself to yield the characteristic g' = 2 signal of the vanadyl (VO2+) cation. Remarkably, however, the protein remains pink, demonstrating a dissociation between color and oxidation state. Guanidinium, in contrast, causes a sizeable red shift in the pink protein's absorption maximum without loss of EPR signal intensity, showing dissociation of color and oxidation state in a complementary way.
Antanaitis
B C
BC
Aisen
P
P
D000135:Acid Phosphatase; D000693:Anaerobiosis; D000818:Animals; D055598:Chemical Phenomena; D002621:Chemistry; D004578:Electron Spin Resonance Spectroscopy; D005260:Female; D007527:Isoenzymes; D008667:Metalloproteins / Q000378:metabolism*; D008982:Molybdenum; D010084:Oxidation-Reduction; D010710:Phosphates / Q000378:metabolism*; D011555:Pseudopregnancy; D013552:Swine; D000071681:Tartrate-Resistant Acid Phosphatase
D007527:Isoenzymes; D008667:Metalloproteins; D010710:Phosphates; D008982:Molybdenum; D000135:Acid Phosphatase; D000071681:Tartrate-Resistant Acid Phosphatase; C022175:ammonium molybdate