Imhof Diana, Nothmann Doreen, Zoda Mohammad Safa, Hampel Kornelia, Wegert Jenny, Böhmer Frank D, Reissmann Siegmund
Institute of Biochemistry and Biophysics, Biological-Pharmaceutical Faculty, Friedrich-Schiller-University, Philosophenweg 12, D-07743 Jena, Germany.
J Pept Sci. 2005 Jul;11(7):390-400. doi: 10.1002/psc.631.
Linear and cyclic phosphopeptides related to the pY2267 binding site of the epithelial receptor tyrosine kinase Ros have been synthesized as ligands for the amino-terminal SH2 (src homology) domain of protein tyrosine phosphatase SHP-1. The synthesis was accomplished by Fmoc-based solid-phase methodology using side-chain unprotected phosphotyrosine for the linear and mono-benzyl protected phosphotyrosine for the cyclic peptides. According to molecular modelling, the incorporation of a glycine residue between Lys (position pY-1 relative to phosphotyrosine) and Asp or Glu (position pY+2) was recommended for the cyclic candidates. The preparation of these peptides was successfully performed by the incorporation of a Fmoc-Xxx(Gly-OAll)-OH (Xxx = Asp, Glu) dipeptide building block that was prepared in solution prior to SPPS. The cyclization was achieved with PyBOP following Alloc/OAll-deprotection. This study demonstrates the usefulness of allyl-type protecting groups for the generation of side-chain cyclized phosphopeptides. Alloc/OAll-deprotection and cyclization are compatible with phosphorylated tyrosine.
已合成与上皮受体酪氨酸激酶Ros的pY2267结合位点相关的线性和环状磷酸肽，作为蛋白酪氨酸磷酸酶SHP-1氨基末端SH2（src同源）结构域的配体。合成采用基于Fmoc的固相方法，线性肽使用侧链未保护的磷酸酪氨酸，环状肽使用单苄基保护的磷酸酪氨酸。根据分子建模，建议在环状候选物的Lys（相对于磷酸酪氨酸的pY-1位置）和Asp或Glu（pY+2位置）之间引入甘氨酸残基。通过引入Fmoc-Xxx(Gly-OAll)-OH（Xxx = Asp、Glu）二肽构建块成功制备了这些肽，该构建块在固相肽合成之前在溶液中制备。在烯丙基酯/烯丙基醚脱保护后，使用PyBOP实现环化。本研究证明了烯丙基型保护基团在生成侧链环化磷酸肽方面的实用性。烯丙基酯/烯丙基醚脱保护和环化与磷酸化酪氨酸兼容。
