Decoding Protein-Protein Interactions through Combinatorial Chemistry: Sequence Specificity of SHP-1, SHP-2, and SHIP SH2 Domains

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• 作者：Michael C. Sweeney ; Anne-Sophie Wavreille ; Junguk Park ; Jonathan P. Butchar ; Susheela Tridandapani ; and Dehua Pei
• 刊名：Biochemistry
• 出版年：2005
• 出版时间：November 15, 2005
• 年：2005
• 卷：44
• 期：45
• 页码：14932 - 14947
• 全文大小：562K
• 年卷期：v.44,no.45(November 15, 2005)
• ISSN：1520-4995

文摘

A general, combinatorial library method for the rapid identification of high-affinity peptideligands of protein modular domains is reported. The validity of this method has been demonstrated bydetermining the sequence specificity of four Src homology 2 (SH2) domains derived from protein tyrosinephosphatase SHP-1 and SHP-2 and inositol phosphatase SHIP. A phosphotyrosyl (pY) peptide librarywas screened against the SH2 domains, and the beads that carry high-affinity ligands of the SH2 domainswere identified and peptides were sequenced by partial Edman degradation and mass spectrometry. Theresults reveal that the N-terminal SH2 domain of SHP-2 is capable of recognizing four different classesof pY peptides. Binding competition studies suggest that the four classes of pY peptides all bind to thesame site on the SH2 domain surface. The C-terminal SH2 domains of SHP-1 and SHP-2 and the SHIPSH2 domain each bind to pY peptides of a single consensus sequence. Database searches using theconsensus sequences identified most of the known as well as many potential interacting proteins of SHP-1and/or SHP-2. Several proteins are found to bind to the SH2 domains of SHP-1 and SHP-2 through anew, nonclassical ITIM motif, (V/I/L)XpY(M/L/F)XP, which corresponds to the class IV peptides selectedfrom the pY library. The combinatorial library method should be generally applicable to other proteindomains.