BIOMAT Database Logo BIOMAT Database Logo

Engineered SH2 Domains for Targeted Phosphoproteomics. 2022

Gregory D Martyn, and Gianluca Veggiani, and Ulrike Kusebauch, and Seamus R Morrone, and Bradley P Yates, and Alex U Singer, and Jiefei Tong, and Noah Manczyk, and Gerald Gish, and Zhi Sun, and Igor Kurinov, and Frank Sicheri, and Michael F Moran, and Robert L Moritz, and Sachdev S Sidhu
Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, 160 College Street, Toronto, Ontario M5S3E1, Canada.

A comprehensive analysis of the phosphoproteome is essential for understanding molecular mechanisms of human diseases. However, current tools used to enrich phosphotyrosine (pTyr) are limited in their applicability and scope. Here, we engineered new superbinder Src-Homology 2 (SH2) domains that enrich diverse sets of pTyr-peptides. We used phage display to select a Fes-SH2 domain variant (superFes; sFes1) with high affinity for pTyr and solved its structure bound to a pTyr-peptide. We performed systematic structure-function analyses of the superbinding mechanisms of sFes1 and superSrc-SH2 (sSrc1), another SH2 superbinder. We grafted the superbinder motifs from sFes1 and sSrc1 into 17 additional SH2 domains and confirmed increased binding affinity for specific pTyr-peptides. Using mass spectrometry (MS), we demonstrated that SH2 superbinders have distinct specificity profiles and superior capabilities to enrich pTyr-peptides. Finally, using combinations of SH2 superbinders as affinity purification (AP) tools we showed that unique subsets of pTyr-peptides can be enriched with unparalleled depth and coverage.

UI MeSH Term Description Entries
D011485 Protein Binding The process in which substances, either endogenous or exogenous, bind to proteins, peptides, enzymes, protein precursors, or allied compounds. Specific protein-binding measures are often used as assays in diagnostic assessments. Plasma Protein Binding Capacity,Binding, Protein
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D013058 Mass Spectrometry An analytical method used in determining the identity of a chemical based on its mass using mass analyzers/mass spectrometers. Mass Spectroscopy,Spectrometry, Mass,Spectroscopy, Mass,Spectrum Analysis, Mass,Analysis, Mass Spectrum,Mass Spectrum Analysis,Analyses, Mass Spectrum,Mass Spectrum Analyses,Spectrum Analyses, Mass
D018909 src Homology Domains Regions of AMINO ACID SEQUENCE similarity in the SRC-FAMILY TYROSINE KINASES that fold into specific functional tertiary structures. The SH1 domain is a CATALYTIC DOMAIN. SH2 and SH3 domains are protein interaction domains. SH2 usually binds PHOSPHOTYROSINE-containing proteins and SH3 interacts with CYTOSKELETAL PROTEINS. SH Domains,SH1 Domain,SH2 Domain,SH3 Domain,src Homology Region 2 Domain,Homology Domain, src,Homology Domains, src,SH Domain,SH1 Domains,SH2 Domains,SH3 Domains,src Homology Domain
D019000 Phosphotyrosine An amino acid that occurs in endogenous proteins. Tyrosine phosphorylation and dephosphorylation plays a role in cellular signal transduction and possibly in cell growth control and carcinogenesis. Tyrosine-O-phosphate,Tyrosine O phosphate
D020543 Proteome The protein complement of an organism coded for by its genome. Proteomes

Related Publications

Gregory D Martyn, and Gianluca Veggiani, and Ulrike Kusebauch, and Seamus R Morrone, and Bradley P Yates, and Alex U Singer, and Jiefei Tong, and Noah Manczyk, and Gerald Gish, and Zhi Sun, and Igor Kurinov, and Frank Sicheri, and Michael F Moran, and Robert L Moritz, and Sachdev S Sidhu
Engineered SH2 domains with tailored specificities and enhanced affinities for phosphoproteome analysis.
February 2019, Protein science : a publication of the Protein Society,
Gregory D Martyn, and Gianluca Veggiani, and Ulrike Kusebauch, and Seamus R Morrone, and Bradley P Yates, and Alex U Singer, and Jiefei Tong, and Noah Manczyk, and Gerald Gish, and Zhi Sun, and Igor Kurinov, and Frank Sicheri, and Michael F Moran, and Robert L Moritz, and Sachdev S Sidhu
ZIP codes for delivering SH2 domains.
January 2004, Cell,
Gregory D Martyn, and Gianluca Veggiani, and Ulrike Kusebauch, and Seamus R Morrone, and Bradley P Yates, and Alex U Singer, and Jiefei Tong, and Noah Manczyk, and Gerald Gish, and Zhi Sun, and Igor Kurinov, and Frank Sicheri, and Michael F Moran, and Robert L Moritz, and Sachdev S Sidhu
SH2 and SH3 domains.
July 1993, Current biology : CB,
Gregory D Martyn, and Gianluca Veggiani, and Ulrike Kusebauch, and Seamus R Morrone, and Bradley P Yates, and Alex U Singer, and Jiefei Tong, and Noah Manczyk, and Gerald Gish, and Zhi Sun, and Igor Kurinov, and Frank Sicheri, and Michael F Moran, and Robert L Moritz, and Sachdev S Sidhu
Lipid Binding of SH2 Domains.
January 2023, Methods in molecular biology (Clifton, N.J.),
Gregory D Martyn, and Gianluca Veggiani, and Ulrike Kusebauch, and Seamus R Morrone, and Bradley P Yates, and Alex U Singer, and Jiefei Tong, and Noah Manczyk, and Gerald Gish, and Zhi Sun, and Igor Kurinov, and Frank Sicheri, and Michael F Moran, and Robert L Moritz, and Sachdev S Sidhu
Molecular recognition by SH2 domains.
January 2002, Advances in protein chemistry,
Gregory D Martyn, and Gianluca Veggiani, and Ulrike Kusebauch, and Seamus R Morrone, and Bradley P Yates, and Alex U Singer, and Jiefei Tong, and Noah Manczyk, and Gerald Gish, and Zhi Sun, and Igor Kurinov, and Frank Sicheri, and Michael F Moran, and Robert L Moritz, and Sachdev S Sidhu
Signalling through SH2 and SH3 domains.
January 1993, Trends in cell biology,
Gregory D Martyn, and Gianluca Veggiani, and Ulrike Kusebauch, and Seamus R Morrone, and Bradley P Yates, and Alex U Singer, and Jiefei Tong, and Noah Manczyk, and Gerald Gish, and Zhi Sun, and Igor Kurinov, and Frank Sicheri, and Michael F Moran, and Robert L Moritz, and Sachdev S Sidhu
Targeting SH2 domains in breast cancer.
January 2014, Future medicinal chemistry,
Gregory D Martyn, and Gianluca Veggiani, and Ulrike Kusebauch, and Seamus R Morrone, and Bradley P Yates, and Alex U Singer, and Jiefei Tong, and Noah Manczyk, and Gerald Gish, and Zhi Sun, and Igor Kurinov, and Frank Sicheri, and Michael F Moran, and Robert L Moritz, and Sachdev S Sidhu
Interactions between SH2 and SH3 domains.
January 1998, Biochemical and biophysical research communications,
Gregory D Martyn, and Gianluca Veggiani, and Ulrike Kusebauch, and Seamus R Morrone, and Bradley P Yates, and Alex U Singer, and Jiefei Tong, and Noah Manczyk, and Gerald Gish, and Zhi Sun, and Igor Kurinov, and Frank Sicheri, and Michael F Moran, and Robert L Moritz, and Sachdev S Sidhu
Functions of SH2 and SH3 domains.
January 1998, Current topics in microbiology and immunology,
Gregory D Martyn, and Gianluca Veggiani, and Ulrike Kusebauch, and Seamus R Morrone, and Bradley P Yates, and Alex U Singer, and Jiefei Tong, and Noah Manczyk, and Gerald Gish, and Zhi Sun, and Igor Kurinov, and Frank Sicheri, and Michael F Moran, and Robert L Moritz, and Sachdev S Sidhu
SH2 domains recognize specific phosphopeptide sequences.
March 1993, Cell,
Copied contents to your clipboard!