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PMID: 20357770 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.

A potent and highly specific FN3 monobody inhibitor of the Abl SH2 domain.

Nature structural & molecular biology ·Vol. 17 ·No. 4 ·2010-04-00 ·Pages 519-27

Wojcik J, Hantschel O, Grebien F, Kaupe I, Bennett KL, Barkinge J, Jones RB, Koide A, Superti-Furga G, Koide S

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.

MeSH Terms
Amino Acid Sequence Antibodies, Monoclonal/immunology Models, Molecular Molecular Sequence Data Oncogene Proteins v-abl/chemistry,immunology Phosphorylation Sequence Homology, Amino Acid src Homology Domains
Chemicals
Antibodies, Monoclonal Oncogene Proteins v-abl
Authors & Affiliations
10 authors, click to expand affiliations / ORCID
Wojcik John
Department of Biochemistry and Molecular Biology, University of Chicago, Chicago, Illinois, USA.
Hantschel Oliver
Grebien Florian
Kaupe Ines
Bennett Keiryn L
Barkinge John
Jones Richard B
Koide Akiko
Superti-Furga Giulio
Koide Shohei
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Article Info
Journal
Nature structural & molecular biology
Abbr.
Nat Struct Mol Biol
ISSN
1545-9985
DOI
10.1038/nsmb.1793
Published
2010-04-00
Epub
2010-00-28
Pages
519-27
Language
English
Region
United States
NLM ID
101186374
PMCID
PMC2926940
Subset
IM
Grants
NIGMS NIH HHS · U54 GM074946 · United States
NIGMS NIH HHS · R01 GM072688-01 · United States
NIGMS NIH HHS · T32 GM007281-33 · United States
NIGMS NIH HHS · T32 GM007281 · United States
NCI NIH HHS · R21-CA132700 · United States
NIGMS NIH HHS · 5T32GM07281-33 · United States
NIGMS NIH HHS · U54-GM74946 · United States
NIGMS NIH HHS · U54 GM074946-01 · United States
NIGMS NIH HHS · R01 GM072688 · United States
NCI NIH HHS · R21 CA132700 · United States
NIGMS NIH HHS · R01-GM72688 · United States
NCI NIH HHS · R21 CA132700-01 · United States
Databases
PDB
3K2M
External Links
PubMed source DOI: 10.1038/nsmb.1793 PMC Full Text View journal details
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