Dataset Information

Peptide ligands that use a novel binding site to target both TGF-? receptors.

ABSTRACT: The transforming growth factor beta (TGF-?) signaling pathway plays myriad roles in development and disease. TGF-? isoforms initiate signaling by organizing their cell surface receptors T?RI and T?RII. Exploration and exploitation of the versatility of TGF-? signaling requires an enhanced understanding of structure-function relationships in this pathway. To this end, small molecule, peptide, and antibody effectors that bind key signaling components would serve as valuable probes. We focused on the extracellular domain of T?R1 (T?RI-ED) as a target for effector screening. The observation that T?RI-ED can bind to a TGF-? coreceptor (endoglin) suggests that the T?RI-ED may have multiple interaction sites. Using phage display, we identified two peptides LTGKNFPMFHRN (Pep1) and MHRMPSFLPTTL (Pep2) that bind the T?RI-ED (K(d)? 10(-5) M). Although our screen focused on T?RI-ED, the hit peptides interact with the T?RII-ED with similar affinities. The peptide ligands occupy the same binding sites on T?RI and T?RII, as demonstrated by their ability to compete with each other for receptor binding. Moreover, neither interferes with TGF-? binding. These results indicate that both T?RI and T?RII possess hot spots for protein-protein interactions that are distinct from those used by their known ligand TGF-?. To convert these compounds into high affinity probes, we exploited the observation that T?RI and T?RII exist as dimers on the cell surface; therefore, we assembled a multivalent ligand. Specifically, we displayed one of our receptor-binding peptides on a dendrimer scaffold. We anticipate that the potent multivalent ligand that resulted can be used to probe the role of receptor assembly in TGF-? function.

SUBMITTER: Li L

PROVIDER: S-EPMC3064480 | biostudies-literature | 2010 Dec

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Peptide ligands that use a novel binding site to target both TGF-β receptors.

Li Lingyin L Orner Brendan P BP Huang Tao T Hinck Andrew P AP Kiessling Laura L LL

Molecular bioSystems 20101004 12

The transforming growth factor beta (TGF-β) signaling pathway plays myriad roles in development and disease. TGF-β isoforms initiate signaling by organizing their cell surface receptors TβRI and TβRII. Exploration and exploitation of the versatility of TGF-β signaling requires an enhanced understanding of structure-function relationships in this pathway. To this end, small molecule, peptide, and antibody effectors that bind key signaling components would serve as valuable probes. We focused on t ...[more]

PMID: 20890540

Similar Datasets

Project description:Homologous to E6AP carboxyl-terminus (HECT)-type E3 ligase performs ubiquitin (Ub)-proteasomal protein degradation via forming a complex with E2∼Ub. Enveloped viruses including SARS-CoV-2 escape from the infected cells by harnessing the E-class vacuolar protein-sorting (ESCRT) machinery and mimic the cellular system through PPAY motif-based linking to HECT Ub ligase activity. In the present study, we have characterized the binding pattern of E2UbcH5B to HECT domains of NEDD4L, WWP1, WWP2, HECW1, and HECW2 through in silico analysis to isolate the E2UbcH5B-specific peptide inhibitors that may target SARS-CoV-2 viral egression. Molecular dynamics analysis revealed more opening of E2UbcH5B-binding pocket upon binding to HECTNEDD4L, HECTWWP1, HECTWWP2, HECTHECW1, and HECTHECW2. We observed similar binding pattern for E2UbcH5B and mentioned HECT domains as previously reported for HECTNEDD4L where Trp762, Trp709, and Trp657 residues of HECTNEDD4L, HECTWWP1, and HECTWWP2 are involved in making contacts with Ser94 residue of E2UbcH5B. Similarly, corresponding to HECTNEDD4L Tyr756 residue, HECTWWP1, HECTWWP2, HECTHECW1, and HECTHECW2-specific Phe703, Phe651, Phe1387, and Phe1353 residues execute interaction with E2UbcH5B. Our analysis suggests that corresponding to Cys942 of HECTNEDD4L, Cys890, Cys838, Cys1574, and Cys1540 residues of HECTWWP1, HECTWWP2, HECTHECW1, and HECTHECW2, respectively are involved in E2-to-E3 Ub transfer. Furthermore, MM-PBSA free energy calculations revealed favorable energy values for E2UbcH5B-HECT complexes along with the individual residue contributions. Subsequently, two E2UbcH5B-derived peptides (His55-Phe69 and Asn81-Ala96) were tested for their binding abilities against HECT domains of NEDD4L, WWP1, WWP2, HECW1, and HECW2. Their binding was validated through substitution of Phe62, Pro65, Ile84, and Cys85 residues into Ala, which revealed an impaired binding, suggesting that the proposed peptide ligands may selectively target E2-HECT binding and Ub-transfer. Collectively, we propose that peptide-driven blocking of E2-to-HECT Ub loading may limit SARS-CoV-2 egression and spread in the host cells.

Dataset Information

Peptide ligands that use a novel binding site to target both TGF-? receptors.

Publications

Peptide ligands that use a novel binding site to target both TGF-β receptors.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets