Summary
Phospholamban (PLB), a 52-residue integral membrane protein of the cardiac sarcoplasmic reticulum, is known as the regulator of the Ca2+-ATPase pump via its phosphorylation-dephosphorylation at Ser16. In order to investigate the structural effects of the phosphorylation on the cytoplasmic domain of PLB, we synthesized PLB 2–33 in its nonphosphorylated and phosphorylated forms using Fmoc chemistry. PLB 2–33 was phosphorylated using the global phosphorylation method. Phosphorylation with di-t-butyl-N,N-diisopropylphosphoramidite led to an incomplete reaction (nonphosphorylated peptide was recovered) and to the formation of an H-phosphonate. In contrast, the use of dibenzyl-N,N-diisopropylphosphoramidite yielded the desired phosphorylated peptide quantitatively and did not give rise to the H-phosphonate. Our results show the effectiveness of the dibenzyl-protected phosphoramidite when the site to be phosphorylated is particularly hindered.
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Kim, H.W., Steenaart, N.A.E., Ferguson, D.G. and Kranias, E.G., J. Biol. Chem., 265 (1990) 1702.
Kitas, E.A., Knorr, R., Trzeciak, A. and Bannwarth, W., Helv. Chim. Acta, 74 (1991) 1314.
Perich, J.W. and Johns, R.B., Tetrahedron Lett., 29 (1988) 2369.
Staerkaer, G., Jakobsen, M.H., Olsen, C.E. and Holm, A., Tetrahedron Lett., 32 (1991) 5389.
Andrews, D.M., Kitchin, J. and Seale, P.W., Int. J. Pept. Protein Res., 38 (1991) 469.
Perich, J.W., LeNguyen, D. and Reynolds, E.C., Tetrahedron Lett., 32 (1991) 4033.
Tegge, W., Int. J. Pept. Protein Res., 43 (1994) 448.
Hoffmann, R., Wachs, W.O., Berger, R.G., Kalbitzer, H.R., Waidelich, D., Bayer, E., Wagner-Redeker, W. and Zeppezauer, M., Int. J. Pept. Protein Res., 45 (1995) 26.
Ottinger, E.A., Shekels, L.L., Bernlohr, D.A. and Barany, G., Biochemistry, 32 (1993) 4354.
Austin, B. and Hawkes, J., In Epton, R. (Ed.) Innovation and Perspectives in Solid Phase Synthesis, Mayflower Worldwide, Birmingham, 1994, pp. 437–440.
Knapp, D.R., Oatis, J.E. and Papac, D.I., Int. J. Pept. Protein Res., 42 (1993) 259.
Terzi, E., Poteur, L. and Trifilieff, E., FEBS Lett., 309 (1992) 413.
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Poteur, L., Trifilieff, E. Global phosphorylation at Ser16 of the 32-residue cytoplasmic domain of phospholamban: Comparison of di-t-butyl- and dibenzyl-N,N-diisopropylphosphoramidites. Lett Pept Sci 2, 271–276 (1996). https://doi.org/10.1007/BF00142238
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DOI: https://doi.org/10.1007/BF00142238