Biopolym. Cell. 1987; 3(5):257-262.
Cell Biology
Interaction of in vitro synthesized casein with artificial phospholipid vesicules
1Balkov D. I., 2Rachkov A. E., 3Kolchinskaya L. I., 1Starodub N. F., 2Elskaya A. V., 1Lishko V. K.
  1. A. V. Palladin Institute of Biochemistry, Academy of Sciences of the Ukrainian SSR
    Kiev, USSR
  2. Institute of Molecular Biology and Genetics, Academy of Sciences of the Ukrainian SSR
    Kiev, USSR
  3. A. A. Bogomoletz Institute of Physiology, Academy of Sciences of the Ukrainian SSR
    Kiev, USSR

Abstract

The model system is developed to study the incorporation of products of eukaryotic mRNA cell-free translation into lecithin liposomes. It is shown that in the presence of casein mRNA the 14C-labeled product of translation is accumulated inside liposomes. When mRNA for globin, a nonsecretory protein, is translated in this cell-free system, 14C-label is not found in the internal volume of liposomes. The polypeptides extracted from liposomes after their incubation in the system of casein mRNA translation interact specifically with anti-casein antibodies. These data provide evidence that the transfer of synthesizing casein through bilayered lipid membrane does not require a specific receptor. Insertion and transfer of this protein occur co-translationally, due to the interaction of «signal peptide» with membrane lipids.

References

[1] Spirin AS. Co-translation folding, compartmentalization and modification of proteins. Mol Biol (Mosk). 1984;18(6):1445-60.
[2] Blobel G, Sabatini DD. Controlled proteolysis of nascent polypeptides in rat liver cell fractions. I. Location of the polypeptides within ribosomes. J Cell Biol. 1970;45(1):130-45.
[3] Sabatini DD, Blobel G. Controlled proteolysis of nascent polypeptides in rat liver cell fractions. II. Location of the polypeptides in rough microsomes. J Cell Biol. 1970;45(1):146-57.
[4] Hortsch M, Meyer DI. Pushing the signal hypothesis: what are the limits? Biol Cell. 1984;52(1 Pt A):1-8.
[5] Adelman MR, Sabatini DD, Blobel G. Ribosome-membrane interaction. Nondestructive disassembly of rat liver rough microsomes into ribosomal and membranous components. J Cell Biol. 1973;56(1):206-29.
[6] Smith WP, Tai PC, Davis BD. Interaction of secreted nascent chains with surrounding membrane in Bacillus subtilis. Proc Natl Acad Sci U S A. 1978;75(12):5922-5.
[7] Ohno-Iwashita Y, Wolfe P, Ito K, Wickner W. Processing of preproteins by liposomes bearing leader peptidase. Biochemistry. 1984;23(25):6178-84.
[8] Meyer DI, Krause E, Dobberstein B. Secretory protein translocation across membranes-the role of the "docking protein'. Nature. 1982;297(5868):647-50.
[9] Walter P, Blobel G. Purification of a membrane-associated protein complex required for protein translocation across the endoplasmic reticulum. Proc Natl Acad Sci U S A. 1980;77(12):7112-6.
[10] Wickner W, Mandel G, Zwizinski C, Bates M, Killick T. Synthesis of phage M13 coat protein and its assembly into membranes in vitro. Proc Natl Acad Sci U S A. 1978;75(4):1754-8.
[11] Nesmeianova MA, Bogdanov MV, Kolot MN, Zemlianukhina OA, Kulaev IS. Interaction of alkaline phosphatase and acid phospholipids in E. coli cells and artificial membranes. Biokhimiia. 1982;47(4):671-7.
[12] Nesmeianova MA. Molecular mechanisms of membrane participation in the biosynthesis of secretory proteins and its regulation in Escherichia coli. Usp Sovrem Biol. 1982;94(2):225-42.
[13] Starodub NF, Rachkov AE, Petik AV et al. Isolation of individual mRNA and immunochemical testing translation products. Methods of molecular biology. Kiev, Naukova dumka, 1986; 90-9.
[14] Limborskaia SA, Frolova LIu, Makarovskaia EE, Khil'ko SN. Isolation of human globin mRNA and synthesis of its complementary DNA by the reverse transcription route. Dokl Akad Nauk SSSR. 1976;230(4):981-4.
[15] Rachkov AE, Starodub NF. Isolation of biologically active mRNA from mammary glands of cows. Molekulyarnaya biologiya. 1984; Is. 37:29-33.
[16] Marcus A, Efron D, Weeks DP. The wheat embryo cell-free system. Methods Enzymol. 1974;30:749-54.
[17] Bangham AD, Hill HW, Miller NGA. Preparation and use of liposomes as model of biological membrane. Meth Membrane Biol. 1974;1:1-68.
[18] Szoka F Jr, Papahadjopoulos D. Procedure for preparation of liposomes with large internal aqueous space and high capture by reverse-phase evaporation. Proc Natl Acad Sci U S A. 1978;75(9):4194-8.
[19] Al-Sarraj K, White DA, Mayer RJ. Purification and properties of casein from mammary gland of lactating rabbits. Int J Biochem. 1978;9(4):269-77.
[20] McConahey PJ, Dixon FJ. Radioiodination of proteins by the use of the chloramine-T method. Methods Enzymol. 1980;70(A):210-3.
[21] Wessel D, Fl?gge UI. A method for the quantitative recovery of protein in dilute solution in the presence of detergents and lipids. Anal Biochem. 1984;138(1):141-3.
[22] Nanba S, Hikawa A, Kitoo N, Horigome T, Omata S, Sugano H. Interaction of secretory protein precursors with phospholipids in liposomes. J Biochem. 1984;96(4):1133-42.
[23] Yoshikawa M, Mizukami T, Omori K, Sasaki R, Chiba H. Isolation and characterization of mRNA from mammary gland of lactating cow. Agr Biol Chem. 1981; 45(1):177-183.
[24] Garnier J, Gaye P, Mercier JC, Robson B. Structural properties of signal peptides and their membrane insertion. Biochimie. 1980;62(4):231-9.