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Abstract
Monoclonal antibodies have been produced against electrophoretically purified MP18, a major calf lens membrane Mr = 18,000 substrate for cAMP-dependent protein kinase. One of these antibodies (monoclonal antibody 2D10) cross-reacted with both native MP18 in lens membranes, and sodium dodecyl sulfate-denatured, electrophoretically purified MP18. In immunoblots, this antibody recognized MP18 in pig, sheep, rat, human, but not chicken lens membranes, indicating the similarity of this protein in mammalian lenses. Amino acid sequencing revealed that the N-terminal sequence of MP18 is identical in these five different mammalian species and is unrelated to any previously sequenced lens or junctional proteins. Electron microscopic examination of monoclonal antibody 2D10-labeled bovine, pig and rat lens membranes indicated that MP18 is localized exclusively to the thicker 16-17 nm junctions in isolated preparations of lens fiber cell membranes. These results provide evidence of a role for MP18 in mammalian lens fiber cell junctional organization.
Highlights
Monoclonal antibodies have been produced against electrophoretically purified MP18, a major calf lens membraneM, = 18,000substrate for CAMP-dependent protein kinase
Electron microscopic examination ofmonoclonal antibody 2D10-labeled bovine, pigand rat lens membranes indicated that MP18 is localized exclusively to the thicker [16,17] nm junctions in isolated preparations of lens fiber cell membranes
Recent studies in our laboratory suggest that MP18 maybe the major lens membrane component which interacts with calmodulin (22)
Summary
F., Klukas, K., Donahue, P., and Johnson, R. (1982) EzEp.ye Res. 3 5 , 113-129 42. J., Robertson, N,, Wong, M.,Zigler,J., and Kinoshita,. (1979) Exp. Eye Res. 2 8 , 359-365 43. Dermietzel, R., Leibstein, A., Frixen, U., Janssen-Timmen, U., Traub, O., and Willecke, K. B., Dermietzel, R., Teplow, D., Traub, O., Willecke, K., and Revel, J.-P. O., Look, J., Dermietzel, R., Brummer, F., Hulser, D., and Willecke, K. K., Traub, O., Look, J., Stutenkemper, R., and Dermietzel, R. E. J., John, S., Kam, E., Meagher, L., andPitts, J. D. (1986) CZBA Symposiumon Functional Complexes of Epithelial Cells (Bok, G., and Clark, S., eds) pp. 92-107, John Wiley, Chichester, England
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