Synthesis of phosphopeptides

JS McMurray, DR Colman IV, W Wang, ML Campbell. The synthesis of phosphopeptides. Biopolymers (Pept Sci) 60, 3-31, 2001. 

Example 24 preparation of an asymmetrically-protected phospho-roamidite 0-tert-butyl-0-2-cyanoethyl-iV,iSr-diisopropylphosphoroamidite as a phosphitylating reagent in the synthesis of phosphopeptides has been described by Toth s group. [52]. This reagent allows the efficient synthesis of serine and threoine containing phosphopeptides by the global approach. 

Synthesis of Phosphopeptides Using the Post-Assembly Phosphorylation Approach... 

The synthesis of phosphopeptides is typically confirmed mass spectrometri-cally using either a MALDI (matrix-assisted laser desorption/ionizafion) or an ESI (electrospray ionization) source, and the peptide purity is determined by reversed-phase chromatography coupled to an UV detector (Figs. 1 and 2B). Whenever possible, phosphopeptide analyses should be complimented by mass spectra recorded in negative ion mode. For most biochemical applications it is necessary to purify the peptides by HPLC techniques (Fig. 2A). 

Wakamiya, T Saruta, K., Yasuoka, J and Kusumoto, S. (1994) An efficient procedure for solid-phase synthesis of phosphopeptides by the Fmoc strategy. Chem. Lett. 1099-1102. 

Enzymatic protecting-group techniques have recently been usedb in the synthesis of phosphopeptides, in which the phenylacetanoide group is removed from the respective phenylacetyl amides by means of PGA. In the course of the ensuing enzymatic transformations only the phenylacetamide-blocking group is attacked. The peptide bonds, the C-terminal ester, and the phosphates remain intact (Scheme 6). More importantly, the reaction conditions are so mild that no trace of P-elimination is observed. ... 

The O-trityl protection can be removed under the same conditions as used for cleavage of peptides from 2-chlorotrityl resin, i.e. with ACOH/TFE/CH2Q2 (2 2 6, rt, 1 h) as the cleavage nnixture or with 1% TFA without affecting tert-butyl side-chain protection. Silanes are added as hydride donors to scavenge the trityl cations and to avoid retritylation. Most commonly, the trityl derivatives of tyrosine, serine, and threonine are used for the synthesis of phosphopeptides by postsynthetic methods on a solid support. In contrast to the use of protected phosphoamino acid derivatives, such as Fmoc-Tyr(POOBzl)-OH,f l both the phosphorylated and nonphosphorylated peptides can be prepared from one syn-... 

Synthesis of phosphopeptides (a) Perich JW (1990) In Kemp BE (ed) Peptides and protein phosphorylation. CRC Press, Boca Raton, p 289 (b) Shapiro G, Buechler D (1994) Tetrahedron Lett 35 5421 (and refs, given therein)... 

Table 2 Reagents for the Synthesis of Phosphopeptides by Phosphorylation on the Solid Support...

T Wakamiya, K Saruta, S Kusumoto. Acid-stable phosphate-protection for phosphoamino acid in the synthesis of phosphopeptides. Proceedings of the Second Japan Symposium on Peptide Chemistry, Shizuoka, 1992, pp 39-41. T Wakamiya, R Togashi, K Saruta, S Kusumoto, S Aimoto, K Yoshizawa-Kumagaye, K Nakajima. New phosphoryl protecting groups for synthesis of phosphopeptides by Boc method. Proceedings of the Thirty-Third Symposium on Peptide Chemistry, Japan, 1995, pp 17-20. 

Folsch, G. Synthesis of phosphopeptides. V. Further dipeptides, tripeptides and O-phosphorylated derivatives of L-serine. Acta Chem. Scand. 20, 459 (1966). 

The following considerations apply to the synthesis of phosphopeptides using Fmoc-Ser(PO(OBzl)OH)-OH, Fmoc-Thr(PO(OBzl)OH)-OH, Fmoc-Tyr-(PO(OBzl)OH)-OH, and Fmoc-Tyr(P03H2)-OH. 

Harris P, Williams G, Shepherd P, Brimble M (2008) The synthesis of phosphopeptides using microwave-assisted solid phase peptide synthesis. Int J Pept Res Ther 14 387-392... 

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