3K2M

Crystal Structure of Monobody HA4/Abl1 SH2 Domain Complex

Summary for 3K2M

• Entry DOI: 10.2210/pdb3k2m/pdb
• Descriptor: Proto-oncogene tyrosine-protein kinase ABL1, Monobody HA4, PHOSPHATE ION, ... (4 entities in total)
• Functional Keywords: engineered binding protein, antibody mimic, protein-protein complex, sh2 domain, atp-binding, phosphoprotein, tyrosine-protein kinase, signaling protein, signaling protein-protein binding complex, signaling protein/protein binding
• Biological source: Homo sapiens (human); More
• Cellular location: Cytoplasm, cytoskeleton. Isoform IB: Nucleus membrane; Lipid-anchor: P00519
• Total number of polymer chains: 4
• Total formula weight: 47015.66

Authors

Wojcik, J.B.,Duguid, E. (deposition date: 2009-09-30, release date: 2010-03-31, Last modification date: 2023-09-06)

Primary citation

Wojcik, J.,Hantschel, O.,Grebien, F.,Kaupe, I.,Bennett, K.L.,Barkinge, J.,Jones, R.B.,Koide, A.,Superti-Furga, G.,Koide, S.
A potent and highly specific FN3 monobody inhibitor of the Abl SH2 domain.
Nat.Struct.Mol.Biol., 17:519-527, 2010

PubMed Abstract: Interactions between Src homology 2 (SH2) domains and phosphotyrosine sites regulate tyrosine kinase signaling networks. Selective perturbation of these interactions is challenging due to the high homology among the 120 human SH2 domains. Using an improved phage-display selection system, we generated a small antibody mimic (or 'monobody'), termed HA4, that bound to the Abelson (Abl) kinase SH2 domain with low nanomolar affinity. SH2 protein microarray analysis and MS of intracellular HA4 interactors showed HA4's specificity, and a crystal structure revealed how this specificity is achieved. HA4 disrupted intramolecular interactions of Abl involving the SH2 domain and potently activated the kinase in vitro. Within cells, HA4 inhibited processive phosphorylation activity of Abl and also inhibited STAT5 activation. This work provides a design guideline for highly specific and potent inhibitors of a protein interaction domain and shows their utility in mechanistic and cellular investigations.

PubMed: 20357770
DOI: 10.1038/nsmb.1793

Experimental method

X-RAY DIFFRACTION (1.75 Å)