PDBsum entry 4je4

Signaling protein/protein binding

PDB id: 4je4

Contents

Protein chains

102 a.a.

93 a.a.

Waters ×85

PDB id:

4je4

Name:

Signaling protein/protein binding

Title:

Crystal structure of monobody nsa1/shp2 n-sh2 domain complex

Structure:

Tyrosine-protein phosphatase non-receptor type 11. Chain: a. Fragment: n-terminal sh2 domain. Synonym: protein-tyrosine phosphatase 1d, ptp-1d, protein-tyrosine phosphatase 2c, ptp-2c, sh-ptp2, shp-2, shp2, sh-ptp3. Engineered: yes. Monobody nsa1. Chain: b. Engineered: yes

Source:

Homo sapiens. Human. Organism_taxid: 9606. Gene: ptp2c, ptpn11, shptp2. Expressed in: escherichia coli. Expression_system_taxid: 469008.

Resolution:

2.31Å R-factor: 0.206 R-free: 0.255

Authors:

F.Sha,S.Koide

Key ref:

F.Sha et al. (2013). Dissection of the BCR-ABL signaling network using highly specific monobody inhibitors to the SHP2 SH2 domains. Proc Natl Acad Sci U S A, 110, 14924-14929. PubMed id: 23980151 DOI: 10.1073/pnas.1303640110

Date:

26-Feb-13 Release date: 28-Aug-13

Protein chain

Q06124  (PTN11_HUMAN) -  Tyrosine-protein phosphatase non-receptor type 11 from Homo sapiens

Seq: 593 a.a.

Struc: 102 a.a.*

Protein chain

P02751  (FINC_HUMAN) -  Fibronectin from Homo sapiens

Seq: 2477 a.a.

Struc: 93 a.a.*

* PDB and UniProt seqs differ at 20 residue positions (black crosses)

Enzyme reactions

• Enzyme class: Chain A: E.C.3.1.3.48 - protein-tyrosine-phosphatase.
• Reaction: O-phospho-L-tyrosyl-[protein] + H2O = L-tyrosyl-[protein] + phosphate

Molecule diagrams generated from .mol files obtained from the KEGG ftp site

Key reference

DOI no: 10.1073/pnas.1303640110

Proc Natl Acad Sci U S A 110:14924-14929 (2013)

PubMed id: 23980151

Dissection of the BCR-ABL signaling network using highly specific monobody inhibitors to the SHP2 SH2 domains.

F.Sha, E.B.Gencer, S.Georgeon, A.Koide, N.Yasui, S.Koide, O.Hantschel.

ABSTRACT

The dysregulated tyrosine kinase BCR-ABL causes chronic myelogenous leukemia in humans and forms a large multiprotein complex that includes the Src-homology 2 (SH2) domain-containing phosphatase 2 (SHP2). The expression of SHP2 is necessary for BCR-ABL-dependent oncogenic transformation, but the precise signaling mechanisms of SHP2 are not well understood. We have developed binding proteins, termed monobodies, for the N- and C-terminal SH2 domains of SHP2. Intracellular expression followed by interactome analysis showed that the monobodies are essentially monospecific to SHP2. Two crystal structures revealed that the monobodies occupy the phosphopeptide-binding sites of the SH2 domains and thus can serve as competitors of SH2-phosphotyrosine interactions. Surprisingly, the segments of both monobodies that bind to the peptide-binding grooves run in the opposite direction to that of canonical phosphotyrosine peptides, which may contribute to their exquisite specificity. When expressed in cells, monobodies targeting the N-SH2 domain disrupted the interaction of SHP2 with its upstream activator, the Grb2-associated binder 2 adaptor protein, suggesting decoupling of SHP2 from the BCR-ABL protein complex. Inhibition of either N-SH2 or C-SH2 was sufficient to inhibit two tyrosine phosphorylation events that are critical for SHP2 catalytic activity and to block ERK activation. In contrast, targeting the N-SH2 or C-SH2 revealed distinct roles of the two SH2 domains in downstream signaling, such as the phosphorylation of paxillin and signal transducer and activator of transcription 5. Our results delineate a hierarchy of function for the SH2 domains of SHP2 and validate monobodies as potent and specific antagonists of protein-protein interactions in cancer cells.