Abstract
α-Lactalbumin (α-LA) is biosynthesized and stored at the smooth endoplasmic reticulum (ER), then transferred to the Golgi lumen when prolactin stimulation of lactose biosynthesis and secretion takes place. Because both environments are composed of membranes, it was of interest to examine the interactions of α-LA with relevant model and biological membranes. Using the ESR spin-labeled fatty acid analog 5-doxyl stearic acid, we found evidence reflecting the insertion of “acid-shocked” molten globule (MG) α-LA into lecithin or phosphatidylserine (PS) multi-lamellar vesicles. An additional ∼3 G immobilization was observed in the α-LA-lecithin sample versus the lipid alone. With PS, the increased immobilization was almost 6 G, reflecting an enhanced effect caused by strong electrostatic interactions between the positively charged protein with the negatively charged headgroup at pH 2.4. This was also reflected in the broadening of the PS:α-LA phase transition. Additionally, we have demonstrated that α-LA in its apo-form also shows similar insertion characteristics with both model and natural lipid membranes. Upon addition of calcium, the apo-form is released from the membrane as the Ca2+-bound protein.
Similar content being viewed by others
References
Acharya, K. R., Ren, J., Stuart, D. I., Phillips, D. C., and Fenna, R. E. (1989). J. Mol. Biol. 208: 99–107.
Acharya, K. R., Ren, J., Stuart, D. I., Phillips, D. C., and Fenna, R. E. (1991). J. Mol. Biol. 221: 571–581.
Banuelos, S., and Muga, A. (1995). J. Biol. Chem. 270: 29910–29915.
Banuelos, S., and Muga, A. (1996). Biochemistry 35: 3892–3898.
Berliner, L. J., and Koga, K. (1987). Biochemistry 26: 3006–3009.
Brew, K., Vanaman, T. C., and Hill, R. L. (1967). J. Biol. Chem. 242: 3747–3749.
Brodbeck, U., and Ebner, K. E. (1976). J. Biol. Chem. 241: 762–764.
Brown, E. M. (1984). J. Dairy Sci. 67: 713–722.
Browne, W. J., North, A. C. T., Phillips, D. C., and Brew, K. (1969). J. Mol. Biol. 42: 65–86.
Cawthern, K. M., Permyakov, E. A., and Berliner, L. J. (1996). Protein Sci. 5: 1394–1405.
Charlwood, J., Hanrahan, S., Tyldesley, R., Langridge, J., Dwek, M., Camilleri, P. (2002). Anal. Biochem. 301: 314–324.
Dolgikh, D. A., Gilmarshov, R. I., Braznikov, E. V., Bychkova, V. O., Semisotnov, G. V., Venjaminov, S. Y., and Ptitsyn, O. B. (1981). FEBS Lett. 136: 311–315.
Dufour, E., Subirade, M., Loupil, F. and Riaublanc, A. (1999). Lait 9, 217–228.
Ebner, K. E. (1970). Accounts Chem. Res. 3: 41–47.
Ebner, K. E. (1973). In Boyer, P. D. (ed.), The Enzymes 9: pp. 363–377.
Grishchenko, V. M., Kalinichenko, L. P., Deikus, G. Y., Veprintsev, D. B., Cawthern, K. M., Berliner, L. J., and Permyakov, E. A. (1996). Biochemistry and Molecular Biology International 38: 453–466.
Halskau, O., Froystein, N. A., Muga, A., Martinez, A. (2002). J. Mol. Biol. 321: 99–110.
Hanssens, I., Houthuys, C., Herremann, W., and VanCauwelaert, F. H. (1980). Biochim. Biophys. Acta 602: 539–557.
Hanssens, I., Herreman, W., and Van Cauwelaert, F. H. (1983). Biochim. Biophys. Acta 728: 291–304.
Hanssens, I., Van Ceunebroeck, J. C., Pottel, H., Preaux, G., and Van Cauwelaert, F. H., (1985). Biochim. Biophys. Acta 817: 154–164.
Herreman, W., Van Tomout, P., Van Cauwelaert, F. H., and Hanssens, I. (1981a). Biochim. Biophys. Acta. 640: 419–429.
Herreman, W., VanCauwelaert, F. H., and Hanssens, I. (1981b). Biochem. International 2: 237–242.
Hill, R. L., and Brew, K. (1975). Adv. Enzymol. Relat. Areas Mol. Biol. 43: 411–490.
Hiraoka, Y., Segawa, T., Kuwajima, K., Sugai, S., and Murai, N. (1980). Biochim. Biophys. Res. Comm. 95: 1098–1104.
Huang, C., and Thompson, T. E. (1970). Meth. Enzymol. 32: 485–489.
Lala, A. K., and Kaul, P. (1992). J. Biol. Chem. 267: 19914–19918.
Lala, A. K., Kaul, P., and Ratnam, P. B. (1995). J. Protein Chem. 14: 601–609.
Marsh, D. (1981). In Membrane Spectroscopy, Springer Verlag, pp. 51–157.
Martin, J., Langer, T., Boteva, R., Schramel, A., Horwich, A. L., and Hartl, F.-U. (1991). Nature 352: 36–42.
McKenzie, H. A., and White, F. H. (1991). Adv. Protein Chem. 41: 173–315.
Mitranic, M. M., and Moscarello, M. A. (1985). Biochim. Biophys. Acta 816: 182–186.
Mitranic, M. M., Boggs, J. M., and Moscarello, M. A. (1983). J. Biol. Chem. 258: 8630–8636.
Mitranic, M. M., Paquet, M. R., and Moscarello, M. A. (1988). Biochim. Biophys. Acta 958: 277–284.
Montich, G. G., and Marsh, D. (1995). Biochemistry 34: 13139–13145.
Mulqueen, P. M., and Kronman, M. J. (1982). Arch. Biochem. Biophys. 215: 28–39.
Murakami, K., Andree, P. J., and Berliner, L. J. (1982). Biochemistry 21: 5488–5494.
Musci, G., and Berliner, L. J. (1985). Biochemistry 24: 3852–3856.
Ohgishi, M., and Wada, A. (1983). FEBS Lett. 164: 21–24.
Ostrovsky, A. V., Kalinichenko, L. P., Emelyanenko, V. I., Klimanov, A. V., and Permyakov, E. A. (1988). Biophys. Chem. 30: 105–115.
Patton, F., and Keenan, T. (1975). Biochim. Biophys. Acta 415: 273–309.
Permyakov, E. A., Kalinichenko, L. P., Morozova, L. A., Yamolenko, V. V., and Burstein, E. A. (1981a). Biochim. Biophys. Res. Comm. 100: 191–197.
Permyakov, E. A., Kalinichenko, L. P., Morozova, L. A., Yamolenko, V. V., and Burstein, E. A. (1981b). Biochim. Biophys. Res. Comm. 102: 1–7.
Permyakov, E. A., Morozova, L. A., and Burstein, E. A. (1985). Biophys. Chem. 21: 21–31.
Reboiras, M. D., and Marsh, D. (1991). Biochim. Biophys. Acta 1053: 259–264.
Rothman, J. E., Miller, R. L., and Urbani, L. J. (1984). J. Cell Biol. 99: 260–271.
Rottem, S., Hubbell, W., Hayflick, L., and McConnell, H. M. (1970). Biochim. Biophys. Acta 219: 104–113.
Stuart, D. I., Acharya, K. R., Walker, N. P. C., Smith, S. G., Lewis, M., and Phillips, D. C. (1986). Nature 324: 84–87.
Rights and permissions
About this article
Cite this article
Chaudhuri, D., Narayan, M. & Berliner, L.J. Conformation-Dependent Interaction of α-Lactalbumin with Model and Biological Membranes: A Spin-Label ESR Study. J Protein Chem 23, 95–101 (2004). https://doi.org/10.1023/B:JOPC.0000016263.50484.e1
Published:
Issue Date:
DOI: https://doi.org/10.1023/B:JOPC.0000016263.50484.e1