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T321P

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

Threonine → Proline at position 321 inside TM1. ClinVar Likely pathogenic. AlphaMissense 0.261 (below threshold) — AM under-call. DynaMut2 ΔΔG +0.13 kcal/mol — essentially neutral. Same position as T321R, proline-introduction at TM1 start.

Interactive 3D Structure

Wild-type reference
Wild-type T321 — hydrogen bond to N325
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DynaMut2 mutant · T321P
Mutant P321 — hydrogen bond to I319 lost (3 contacts lost)
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Bond changes · DynaMut2 interaction analysis

3 lost2 gained3 preserved
Interaction typeWild-type partnerMutant partnerStatus
Hydrogen bondI319Lost
Hydrogen bondN325N325Preserved
Polar contactI319I319Preserved
Polar contactI324Lost
Polar contactN325N325Preserved
Van der WaalsI319Gained
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
0.13kcal/mol
Stabilising — mild
AlphaMissense
0.261
LBen
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)Ultra-rare · AF 0.00021%
cDNA changec.961A>C
ClinVar accessionVCV000918064
Last evaluated2025/03/10 00:00

Observed at very low frequency in gnomAD.

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)
AF 0.00021% · 3 / 1,461,866 alleles
Homozygotes
0
Highest-frequency population
European (non-Finnish) · AF 0.00027%

Highest in European (non-Finnish): AF 0.00027% (3 of 1,111,994 alleles), in line with the global figure.

No homozygotes reported in gnomAD v4. For a recessive allele this is expected and is not, on its own, evidence either way.

Ancestry groupAllele freq.AC / ANHom.Carrier est.
European (non-Finnish)0.00027%3 / 1,111,9940~1 in 185330

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 at the start of TM1. Same neighbor environment as T321R: HIS322 (2.5 Å), PRO320 (2.5 Å — adjacent proline!), ILE319 (3.8 Å), ILE324 (4.4 Å), HIS323 (4.5 Å).

Replacing T321 with proline creates a Pro-Pro motif (P320-P321) in the cytoplasmic-to-TM1 boundary region. Two adjacent prolines produce a rigid backbone segment with restricted conformational options — similar to the L723P/P724S Pro-Pro region in the lumenal domain.

The ΔΔG of +0.13 (essentially neutral) reflects fold accommodation through local rearrangement. AlphaMissense's 0.261 below threshold is AM under-call. ClinVar Likely Pathogenic establishes clinical relevance.

Amino-acid chemistry
Threonine (T) → Proline (P) — small polar hydroxyl replaced by rigid helix-breaking residue.
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 = +0.13 — fold essentially unchanged. AlphaMissense 0.261 below threshold.

Mechanism is creation of a P320-P321 Pro-Pro motif at the TM1 boundary, altering TM1 insertion geometry. Therapeutic strategy: site-directed at TM1 start.

Why this matters

T321P + T321R both at position 321, both AM under-calls, both ClinVar pathogenic. The dense TM1 variant cluster (W314R, H313Y, T321R, T321P, H323R, A326E) makes TM1 a high-confidence multi-variant therapeutic target.
Therapeutic Strategy Handoff · prediction

Feed this card to Wolfram Intelligence

Download the T321P 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 T321P PDF card ↓Strategies are AI-generated predictions, not validated therapeutics.

UniProt Domain Annotation

Chain1890 · Wolframin
Transmembrane314334 · Helical
Region1321 · Interaction with ATP6V1A