8FG1

Human diaphanous inhibitory domain bound to diaphanous autoregulatory domain

Summary for 8FG1

• Entry DOI: 10.2210/pdb8fg1/pdb
• NMR Information: BMRB: 31064
• Descriptor: Protein diaphanous homolog 1 (2 entities in total)
• Functional Keywords: inhibitory domain, autoregulatory domain, protein binding
• Biological source: Homo sapiens (human); More
• Total number of polymer chains: 2
• Total formula weight: 32277.92

Authors

Ramirez, L.M.S.,Theophall, G.,Premo, A.,Manigrasso, M.,Yepuri, G.,Burz, D.,Ramasamy, R.,Schmidt, A.M.,Shekhtman, A. (deposition date: 2022-12-12, release date: 2023-10-25, Last modification date: 2024-05-15)

Primary citation

Theophall, G.G.,Ramirez, L.M.S.,Premo, A.,Reverdatto, S.,Manigrasso, M.B.,Yepuri, G.,Burz, D.S.,Ramasamy, R.,Schmidt, A.M.,Shekhtman, A.
Disruption of the productive encounter complex results in dysregulation of DIAPH1 activity.
J.Biol.Chem., 299:105342-105342, 2023

PubMed Abstract: The diaphanous-related formin, Diaphanous 1 (DIAPH1), is required for the assembly of Filamentous (F)-actin structures. DIAPH1 is an intracellular effector of the receptor for advanced glycation end products (RAGE) and contributes to RAGE signaling and effects such as increased cell migration upon RAGE stimulation. Mutations in DIAPH1, including those in the basic "RRKR" motif of its autoregulatory domain, diaphanous autoinhibitory domain (DAD), are implicated in hearing loss, macrothrombocytopenia, and cardiovascular diseases. The solution structure of the complex between the N-terminal inhibitory domain, DID, and the C-terminal DAD, resolved by NMR spectroscopy shows only transient interactions between DID and the basic motif of DAD, resembling those found in encounter complexes. Cross-linking studies placed the RRKR motif into the negatively charged cavity of DID. Neutralizing the cavity resulted in a 5-fold decrease in the binding affinity and 4-fold decrease in the association rate constant of DAD for DID, indicating that the RRKR interactions with DID form a productive encounter complex. A DIAPH1 mutant containing a neutralized RRKR binding cavity shows excessive colocalization with actin and is unresponsive to RAGE stimulation. This is the first demonstration of a specific alteration of the surfaces responsible for productive encounter complexation with implications for human pathology.

PubMed: 37832872
DOI: 10.1016/j.jbc.2023.105342

Experimental method

SOLUTION NMR