Iron content and molecular weight of uteroferrin and a comparison of its iron and copper forms.
The molecular weight of the purple, iron-containing glycoprotein uteroferrin has been determined to be 35140 +/- 1610 by sedimentation equilibrium centrifugation. This is consistent with measurements carried out by other procedures. The dry weight of a solution of uteroferrin with an absorption of 1.00 cm -1 at 280 nm was 0.87 mg/ml. Highly purified uteroferrin has a ratio of absorbance at 280 nm to 545 nm of about 13.2 and a revised extinction coefficient at 545 nm of 3.1 . 10(3) M -1 is presented. The iron content of uteroferrin has been determined both colorimetrically and by atomic absorption spectroscopy. One iron atom is present per polypeptide chain. Reduction with dithionite leads to loss of iron, allowing the apoprotein to be prepared. Enzymatic activity can be restored to the apoprotein with Fe(III) or with Cu(II). The copper enzyme has no visible color and has a higher pH optimum than the Fe-uteroferrin. Whereas the phosphatase activity of the latter is increased several-fold by beta-mercaptoethanol, reduction inactivates Cu-uteroferrin. Both forms of uteroferrin have similar Km values for p--nitrophenyl phosphate and are inhibited by molybdate but not by tartrate. Excess Cu(II) and Fe(III) strongly inhibit uteroferrin phosphatase activity and these results may explain the failure of other to restore activity to the apoprotein using Cu(II) and Fe(III) salts.
D000135:Acid Phosphatase; D000818:Animals; D001059:Apoproteins / Q000378:metabolism; D003300:Copper / Q000494:pharmacology; D019327:Copper Sulfate; D004227:Dithionite / Q000494:pharmacology; D005260:Female; D005290:Ferric Compounds / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007501:Iron / Q000032:analysis*; D007527:Isoenzymes; D008667:Metalloproteins / Q000032:analysis / Q000378:metabolism*; D008970:Molecular Weight; D009596:Nitrophenols / Q000378:metabolism; D009943:Organophosphorus Compounds / Q000378:metabolism; D010084:Oxidation-Reduction; D011247:Pregnancy; D013057:Spectrum Analysis; D013552:Swine; D000071681:Tartrate-Resistant Acid Phosphatase
D001059:Apoproteins; D005290:Ferric Compounds; D007527:Isoenzymes; D008667:Metalloproteins; D009596:Nitrophenols; D009943:Organophosphorus Compounds; D004227:Dithionite; C008644:nitrophenylphosphate; D003300:Copper; D007501:Iron; D000135:Acid Phosphatase; D000071681:Tartrate-Resistant Acid Phosphatase; D019327:Copper Sulfate