No alpha-lactalbumin-like activity detected in a low molecular mass protein fraction of rat epididymal extract.
The same low M(r) protein fraction of epididymal luminal fluid as that studied previously by Hamilton in 1981 was assayed for alpha-lactalbumin (alpha-lac) activity with bovine milk galactosyltransferase (GalTase) as the enzyme source and bovine serum albumin (BSA) to make the total protein concentrations of all the samples in each assay constant. No alpha-lac activity could be detected in this fraction. When glucose was used as an acceptor, radioactivity passing through the ion exchange column used to retain the remaining UDP-galactose did increase with the amount of the proteins of the low M(r) fraction, but this increase was observed not only for samples with an acceptor but also for samples without. The increased radioactive product co-migrated in high-voltage electrophoresis with galactose, not lactose. When GlcNAc was used as an acceptor, the situation was the same, and the increased radioactive product co-migrated with galactose, not LacNAc. No inhibition of bovine milk GalTase activity by the low M(r) proteins was observed. Addition of protein, either BSA or the low M(r) fraction of epididymal luminal fluid, to assay medium that contained no proteins other than GalTase enhanced the GalTase activity both in forming lactose in the presence of glucose and also LacNAc in the presence of GlcNAc. It appears that the earlier reports of alpha-lac-like activity in epididymal fluids and extracts may have been due to the presence of enzymes liberating free galactose from UDP-galactose and/or a stimulatory non-specific effect of the protein in the solutions on the lactose synthesis activity of the GalTase.
Tang
Y
Y
Department of Animal Sciences, University of Adelaide, Waite Agricultural Research Institute, Glen Osmond, Australia.
D000818:Animals; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D004590:Electrophoresis, Paper; D004591:Electrophoresis, Polyacrylamide Gel; D004822:Epididymis / Q000737:chemistry*; D007768:Lactalbumin / Q000032:analysis* / Q000302:isolation & purification; D007785:Lactose / Q000096:biosynthesis; D008297:Male; D051381:Rats; D017208:Rats, Wistar
D007768:Lactalbumin; D007785:Lactose