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The structural integrity of human TFF1 under reducing conditions
Dilsah Nur Elmaci1, Gene Hopping2, Werner Hoffmann3
1Molecular Simulations and Design Group, Max Planck Institute for Dynamics of Complex Technical Systems, 39106 Magdeburg, Germany.
Redox Biology
|February 20, 2025
Summary
The human trefoil factor family 1 (TFF1) domain shows remarkable stability, resisting reduction of its disulfide bonds even under harsh conditions. This resilience ensures TFF1
Area of Science:
- Biochemistry
- Structural Biology
- Molecular Dynamics
Background:
- Trefoil factor family (TFF) peptides, including TFF1, are crucial for gastrointestinal mucosal integrity.
- The TFF domain's structure is stabilized by three intramolecular disulfide bonds formed by six conserved cysteine residues.
Purpose of the Study:
- To investigate the stability of the human TFF1 domain under reducing conditions.
- To understand the resistance of TFF1 disulfide bonds to reduction by tris(2-carboxyethyl)phosphine (TCEP).
Main Methods:
- Experimental investigation of TFF1 domain stability using varying concentrations of TCEP.
- All-atom molecular dynamics simulations (24 μs) of TFF1 under different disulfide bond reduction states.
- Comparison of TFF1 stability with reference peptides like BPTI and linaclotide.
Main Results:
- TFF1 disulfide bonds exhibit high resistance to TCEP reduction compared to BPTI and linaclotide.
- Complete reduction of TFF1 requires a large TCEP excess (150-fold), with no partially reduced intermediates observed.
- Molecular dynamics simulations confirm TFF1 domain compactness and stability, with reduced cysteines remaining buried and non-covalent interactions compensating for lost disulfide bonds.
Conclusions:
- The human TFF1 domain possesses exceptional stability under reducing conditions due to persistent non-covalent interactions.
- This inherent stability supports TFF1's functional resilience during its expression and secretion throughout the body.
- Findings explain the inability to achieve partial reduction and alkylation of TFF1 in experiments.
