Purification of a 240 kDa protein from serum and follicular fluid of water buffalo and its identification as haptoglobin.

The fluids from healthy growing follicles of water buffalo were previously found free of the polypeptides H (M(r) 36,000) and L (M(r) 21,000) which were instead detected in fluids from atretic follicles and blood. Here we report evidence that these two polypeptides, as selected from serum by specific anti-L antibodies, are the subunits of an oligomeric protein. The protein was purified from serum or follicular fluid, and its molecular weight (240 kDa), isoelectric point (6.5), and amino acid composition were determined. The NH2-terminal sequences of the subunits L and H were analyzed: 100% and 90% homology with alpha and beta chains of bovine haptoglobin, respectively, was found. Thus, haptoglobin can be used as a novel molecular marker to assess the physiological state of the blood-follicle barrier or discriminate between atretic and healthy follicles.