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T321R

Category 4 — Stable Fold, Function DisruptedLikely pathogenicTransmembrane · predictedEditorial
ThreonineArginine at position 321 · TM1 (314-334), helical transmembrane · WFS1 (Wolframin)

Threonine → Arginine at position 321 inside TM1. ClinVar Likely pathogenic. AlphaMissense 0.449 (below threshold) — AM under-call. DynaMut2 ΔΔG -0.08 kcal/mol — essentially neutral.

Interactive 3D Structure

Wild-type reference
Wild-type T321 — hydrogen bond to N325
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DynaMut2 mutant · T321R
Mutant R321 — hydrogen bond to I319 lost (2 contacts lost)
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Bond changes · DynaMut2 interaction analysis

2 lost1 gained4 preserved
Interaction typeWild-type partnerMutant partnerStatus
Hydrogen bondI319I319Preserved
Hydrogen bondN325N325Preserved
Polar contactI319I319Preserved
Polar contactI324Lost
Polar contactN325N325Preserved
Van der WaalsN325Gained
HydrophobicI319Lost

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
0.449
Amb
AlphaFold pLDDT
75
model confidence
Schema
Cat 4
Category 4 — Stable Fold, Function Disrupted
ΔΔG confidence: Reduced confidence · Transmembrane
  • transmembrane position; predictor benchmarked on soluble proteins

This variant sits in the transmembrane band. The stability value was computed with a predictor benchmarked on water-soluble proteins. Published benchmarks of ddG predictors on membrane proteins report correlation below 0.4. Treat the magnitude as unreliable and the sign as weak evidence only.

Do not rank this value against a variant from a different region on ΔΔG alone.

Clinical Evidence

ClinVar classificationLikely pathogenic
Review statuscriteria provided, single submitter
Associated conditions(no specific conditions catalogued)
InheritanceInheritance not specified.
Population frequency (gnomAD v4)Absent from gnomAD v4
cDNA changec.962C>G
ClinVar accessionVCV001481097
Last evaluated2025/08/12 00:00

Not observed in ~730k individuals — consistent with a rare allele (ACMG PM2_supporting).

Classified for1★ unverified submission

ClinVar classifies this variant as Likely pathogenic, but does not state what it is classified for. ClinVar records the condition only as "not provided". A classification without a phenotype and an inheritance mode cannot be read as a statement about Wolfram syndrome risk or severity.

  • Phenotype scope is interpretation of submitted records. The computed values on this card (AlphaMissense, DynaMut2 ΔΔG, pLDDT) are measurements of the model, not of a patient.
  • Presentation cannot be generalised from neighbouring variants: one nucleotide over can present very differently. Structural similarity supports a mechanistic hypothesis, never a phenotype claim.

Review status: 1★ single submitter. ClinVar "conditions" is the union of all submissions; 0–1★ entries are submitted conditions, not documented phenotypes.

Population frequency
Global (all ancestries)
Absent from gnomAD v4
Homozygotes
Not available

No population breakdown — the variant is absent from gnomAD v4, so no ancestry group has an observed frequency.

No homozygotes — the variant is absent from gnomAD v4 altogether.

Source: gnomAD r4, retrieved 2026-08-21. Values are allele frequencies. The carrier estimate is derived from the allele frequency (Hardy-Weinberg, ~2·AF) and is an approximation, not a published figure — where a published carrier frequency exists it outranks this column and is cited on the card.

Structural Context

Position 321 sits in TM1, same neighbor environment as T321P (Atlas card adjacent): HIS322 (2.5 Å — partner of H323R and A326E Atlas cards through the H322-H323-A326 cluster), PRO320 (2.5 Å), ILE319 (3.8 Å), ILE324 (4.4 Å), HIS323 (4.5 Å).

Replacing T321 with arginine introduces a large positive charge into the bilayer-embedded TM1. The arginine side chain likely extends toward the membrane-water interface. The H322-H323 cluster nearby is affected by the new positive charge.

The |ΔΔG| of 0.08 (essentially zero) indicates fold accommodates the substitution. AlphaMissense's 0.449 is below threshold — AM under-call. ClinVar Likely Pathogenic establishes clinical relevance. T321R + T321P (same position, different chemistries) both pathogenic confirm position 321 as functionally important.

Amino-acid chemistry
Threonine (T) → Arginine (R) — small polar hydroxyl replaced by large positively-charged guanidinium. Charge introduction into the bilayer.
Position in the protein
TM1 (residues 314–334) · position 321 near the start of TM1 (pLDDT 75).

Druggability Assessment

Category 4 — Stable Fold, Function Disrupted (AM under-call). |ΔΔG| = 0.08 — fold unchanged. AlphaMissense 0.449 below threshold.

Mechanism is charge introduction into TM1. Therapeutic strategy: TM1 microregion site-directed.

Why this matters

T321R + T321P together establish position 321 as a TM1 pathogenic hotspot. Both AM under-calls; both ClinVar pathogenic. Drug discovery here pauses for wet-lab work.
Therapeutic Strategy Handoff · prediction

Feed this card to Wolfram Intelligence

Download the T321R 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 T321R PDF card ↓Strategies are AI-generated predictions, not validated therapeutics.

UniProt Domain Annotation

Chain1890 · Wolframin
Transmembrane314334 · Helical
Region1321 · Interaction with ATP6V1A