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T641K

Category 3/4 — Most DruggableConflictingTransmembrane · predictedEditorial
ThreonineLysine at position 641 · TM10 (632-652), helical transmembrane · WFS1 (Wolframin)

Threonine → Lysine at position 641 inside wolframin's tenth transmembrane helix (TM10). ClinVar carries conflicting classifications — pathogenicity is documented but not universally confirmed. AlphaMissense 0.980, DynaMut2 ΔΔG -0.08 kcal/mol (essentially no destabilization). A near-zero-ΔΔG pathogenic variant with a striking mechanism.

Interactive 3D Structure

Wild-type reference
Wild-type T641 — hydrogen bond to L637
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DynaMut2 mutant · T641K
Mutant K641 — hydrogen bond to V638 lost (2 contacts lost)
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Bond changes · DynaMut2 interaction analysis

2 lost0 gained8 preserved
Interaction typeWild-type partnerMutant partnerStatus
Hydrogen bondL637L637Preserved
Hydrogen bondV638V638Preserved
Hydrogen bondL645L645Preserved
Polar contactL637L637Preserved
Polar contactV638Lost
Polar contactV644V644Preserved
Polar contactL645L645Preserved
Van der WaalsL637L637Preserved
Van der WaalsL645Lost
HydrophobicF414F414Preserved

Lost / gained / preserved interatomic contacts at the variant residue, from the DynaMut2 (Arpeggio) interaction analysis of the wild-type and energy-minimized mutant structures.

Computational Predictions

DynaMut2 ΔΔG
-0.08kcal/mol
Destabilising — mild
AlphaMissense
0.980
LPath
AlphaFold pLDDT
87
model confidence
Schema
Cat 3/4
Category 3/4 — Most Druggable

Clinical Evidence

ClinVar classificationConflicting classifications of pathogenicity
Review statuscriteria provided, conflicting classifications
Associated conditionsWolfram syndrome 1; Wolfram-like syndrome
InheritanceDocumented in association with both Wolfram syndrome 1 (AR) and Wolfram-like syndrome (AD). ClinVar's conflicting classifications reflect that the variant's functional consequences depend on context — homozygous, compound heterozygous, or heterozygous with different penetrance.
Population frequency (gnomAD v4)Ultra-rare · AF 0.00093%
cDNA changec.1922C>A
ClinVar accessionVCV000166599
Last evaluated2025/02/27 00:00

Observed at very low frequency in gnomAD.

Structural Context

Position 641 sits inside TM10, one of wolframin's eleven transmembrane helices anchoring the protein in the ER membrane. The AlphaFold model places T641 within 5 Å of immediate sequence neighbors LEU640 (2.5 Å) and ALA642 (2.5 Å), and into a hydrophobic cluster with LEU637 (3.8 Å), VAL638 (4.0 Å), PHE414 (4.1 Å, from TM3 — indicating helix-helix contact), and ILE643 (4.5 Å). The wild-type threonine's small polar character fits well in this membrane-embedded helix — its hydroxyl can participate in a localized hydrogen bond pattern within the helix backbone.

Replacing threonine with lysine in this position is striking. Lysine's positively-charged primary amine and its long alkyl chain are exceptionally costly in a transmembrane context: charge buried in the bilayer hydrophobic core is thermodynamically unfavorable, and the larger volume cannot be accommodated without local rearrangement. Yet DynaMut2 returns a |ΔΔG| of only 0.08 kcal/mol — essentially no destabilization.

The explanation is geometric. The lysine side chain, despite being large and charged, is flexible and can extend outward toward the membrane-water interface where the charge can be partially satisfied by interaction with lipid headgroups or water. The fold can absorb the substitution. But — and this is the mechanistic insight — the functional integrity of TM10 depends on its packing against TM3 (the PHE414 contact at 4.1 Å). Introducing a charge into that interface, even if the fold accommodates it, disrupts the helix-helix geometry that the wild-type relied on. The result is a variant the protein can fold but cannot function correctly.

AlphaMissense's score of 0.980 reflects this functional severity even though the structural cost is near-zero. The variant is pathogenic by mechanism, not by misfolding.

Amino-acid chemistry
Threonine (T) → Lysine (K) — a small polar residue with a hydroxyl group replaced by a large, positively charged residue with a flexible alkyl chain and a primary amine. The chemistry shift inside a transmembrane helix is unusually severe.
Position in the protein
TM10 (residues 632–652) · position 641 is bilayer-embedded, inside a helical transmembrane segment of wolframin. The lipid environment penalizes charged side chains heavily.

Druggability Assessment

Category 4 — Stable Fold, Function Disrupted. |ΔΔG| = 0.08 kcal/mol — the fold is essentially unperturbed. But the variant is pathogenic by AlphaMissense and clinical evidence, which means the mechanism is functional disruption rather than fold instability.

The most likely mechanism is disrupted TM3-TM10 helix-helix packing: the introduced charge at the helix-helix interface (4.1 Å from PHE414 in TM3) is unfavorable in the bilayer hydrophobic core and would perturb the relative geometry of the two helices. The therapeutic strategy is site-specific: a small molecule that stabilizes the TM3-TM10 packing interface, occupying the geometric niche the wild-type threonine maintained.

This is a variant the Atlas captures particularly well — pre-atlas, the near-zero ΔΔG might have led screeners to deprioritize this position. The atlas tells you which interface to target.

Why this matters

T641K illustrates why fold stability alone is not sufficient to identify therapeutic targets. The protein folds, the variant is pathogenic, and the mechanism is helix-helix interface disruption. Drug discovery here aims at the TM3-TM10 interface, not at the TM10 helix itself. The Atlas's neighbor analysis — surfacing the PHE414 contact across helices — is what makes that target visible.
Therapeutic Strategy Handoff · prediction

Feed this card to Wolfram Intelligence

Download the T641K PDF below and upload it to Wolfram Intelligence to generate therapeutic-strategy proposals — guanidinium mimetics, sigma-1 agonist docking, NAC thiol-capping. NAC is already on the bench-testing list.

Download T641K PDF card ↓Strategies are AI-generated predictions, not validated therapeutics.

UniProt Domain Annotation

Chain1890 · Wolframin
Transmembrane632652 · Helical