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Characterization of lactoferrin binding to the MAC-T bovine mammary epithelial cell line using a biotin-avidin
J J Rejman1, J D Turner, S P Oliver
1Department of Animal Science, University of Tennessee, Knoxville 37901-1071.
Abstract:
1. A non-radioisotopic method utilizing a biotin-avidin approach was used to characterize lactoferrin binding to the clonal MAC-T bovine mammary epithelial cell line. 2. Binding of lactoferrin to MAC-T cells and isolated membranes was specific and saturable. 3. Unlabeled lactoferrin competed for and displaced biotin-labeled lactoferrin from binding sites on mammary epithelial cells. In contrast, unlabeled transferrin did not compete. 4. Scatchard analysis of lactoferrin binding to MAC-T cell crude membranes was nonlinear, revealing two classes of binding sites with association constants (Ka) of 2.36 x 10(7) and 3.36 x 10(6) M-1. 5. Binding of lactoferrin to MAC-T cells may be associated with the initial events which result in decreased MAC-T cell proliferation.