Isolation and characterization of a high molecular weight stable pink form of uteroferrin from uterine secretions and allantoic fluid of pigs.
A pink, high molecular weight form of uteroferrin (Uf) has been isolated from uterine secretions and allantoic fluid of pigs. This protein fraction (denoted FIII) which is relatively stable under physiological conditions of pH, ionic strength, and temperature has a molecular weight of about 80,000, a value approximately twice that of purple Uf (Mr approximately 35,000) isolated from a separate fraction (FIV) by gel filtration. The visible absorption spectrum, EPR signal, and acid phosphatase activity of Uf in FIII are almost identical to those of FIV Uf after the latter has been reduced by 2-mercaptoethanol. However, unlike reduced FIV Uf, the pink, high molecular form does not revert to purple, nor does it show loss of EPR signal and phosphatase activity in the presence of oxygen. In addition, it does not become purple at orthophosphate concentrations which inhibit Uf acid phosphatase activity. Polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate has shown that FIII consists of approximately equal amounts of Uf polypeptides (Mr = 35,000 and 37,000) and a group of three polypeptides (Mr = 40,000, 46,000, and 50,000) antigenically unrelated to Uf. The latter share a common epitope not found on Uf and are probably differentially processed forms of the same protein. FIII can be dissociated by pH conditions below 5.0, by exposure to antibodies raised against Uf or the associated polypeptides, and by sodium dodecyl sulfate at 100 degrees C. The polypeptides in FIII are not therefore linked by disulfide bonds. Treatment with dimethyl suberimidate, however, results in a cross-linked complex (Mr approximately 82,000) consisting of Uf and the associated polypeptides. It is concluded that this high Mr form of Uf is a heterodimer of fully activated Uf and a second polypeptide of unknown function.
D000135:Acid Phosphatase / Q000378:metabolism; D000482:Allantois / Q000378:metabolism*; D000818:Animals; D001667:Binding, Competitive; D003429:Cross Reactions; D004120:Dimethyl Suberimidate / Q000494:pharmacology; D004578:Electron Spin Resonance Spectroscopy; D005321:Extraembryonic Membranes / Q000378:metabolism*; D005260:Female; D006863:Hydrogen-Ion Concentration; D007527:Isoenzymes; D008667:Metalloproteins / Q000302:isolation & purification* / Q000378:metabolism; D008970:Molecular Weight; D009994:Osmolar Concentration; D011247:Pregnancy; D013552:Swine; D000071681:Tartrate-Resistant Acid Phosphatase; D013696:Temperature; D014599:Uterus / Q000378:metabolism*
D007527:Isoenzymes; D008667:Metalloproteins; D004120:Dimethyl Suberimidate; D000135:Acid Phosphatase; D000071681:Tartrate-Resistant Acid Phosphatase