# WFS1 Wolframin — T321R Variant Card

**Molecular Atlas Pilot · RareResearch.AI · Generated by wolfram-variant-card skill**

*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.*

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## Identity

| Field | Value |
|---|---|
| **Variant** | T321R (p.Threonine321Arginine) |
| **DNA change** | c.962C>G |
| **Gene · Protein** | WFS1 · Wolframin (890 aa) |
| **UniProt** | O76024 · WFS1_HUMAN |
| **ClinVar accession** | VCV001481097 |
| **Amino acid change** | Threonine (T) → Arginine (R) — small polar hydroxyl replaced by large positively-charged guanidinium. Charge introduction into the bilayer. |

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## Structural Context

| Field | Value |
|---|---|
| **AlphaFold model** | AF-O76024-F1, v6 |
| **pLDDT at residue 321** | **75.12** — well-folded |
| **Domain** | TM1 (314-334), helical transmembrane |
| **Position context** | TM1 (residues 314–334) · position 321 near the start of TM1 (pLDDT 75). |
| **IDR flag** | No — pLDDT above 50 threshold |

**UniProt features at this position:**

  - Chain: 1-890 Wolframin
  - Transmembrane: 314-334 Helical
  - Region: 1-321 Interaction with ATP6V1A

> 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.

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## Computational Predictions

### AlphaMissense
| Field | Value |
|---|---|
| am_pathogenicity | **0.4487** |
| am_class | **Amb** |
| Interpretation | Likely benign (threshold 0.564) |

### DynaMut2
| Field | Value |
|---|---|
| ΔΔG (kcal/mol) | **-0.08 (Destabilising)** |
| Job ID | 177992011202 |
| Result URL | https://biosig.lab.uq.edu.au/dynamut2/results_prediction/177992011202 |

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## Clinical Evidence

| Field | Value |
|---|---|
| Classification | **Likely pathogenic** |
| Review status | criteria provided, single submitter |
| Last evaluated | 2025/08/12 00:00 |
| Inheritance | Inheritance not specified. |
| WFS1 variant landscape | T321R is 1 of ~326 pathogenic-spectrum variants in WFS1 (out of 2,243 catalogued in ClinVar) |

- (no specific conditions catalogued)

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## Research Path Decision Tree

```
ΔΔG < 2  + binding site affected   →  CATEGORY 3 — docking experiments
ΔΔG 2–4                            →  CATEGORY 2 — pharmacological chaperones
ΔΔG > 4                            →  CATEGORY 1 — gene therapy
pLDDT < 50                         →  CATEGORY 5 — IDR, experimental only
Stable fold + functional site hit  →  CATEGORY 4 — site-specific docking
```

## Final Schema Categorization

**Category 4 — Stable Fold, Function Disrupted**

<strong>Category 4 — Stable Fold, Function Disrupted (AM under-call).</strong> |ΔΔG| = 0.08 — fold unchanged. AlphaMissense 0.449 below threshold.<br/><br/>Mechanism is charge introduction into TM1. Therapeutic strategy: TM1 microregion site-directed.

**Why this card 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.

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## Files in this folder

- `AF-O76024-F1-model_v6.pdb` — AlphaFold structure
- `T321R_molstar_viewer.html` — interactive 3D viewer (auto-highlights position 321 with ball-and-stick + neighbors within 5Å)
- `T321R_variant_card.md` — this card (source of truth)
- `T321R_variant_card.html` — styled printable card
- `T321R_dynamut2_summary.html` — clean offline DynaMut2 result card
- `dynamut2_result.json` — structured result data
- `dynamut2_result_page.html` — local snapshot of the Biosig result page (asset URLs absolutized)
- `T321R_wildtype_interactions.pse` / `T321R_mutant_interactions.pse` — PyMOL sessions

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*Generated by wolfram-variant-card skill · RareResearch.AI Molecular Atlas*
*Every assumption documented. Every score sourced.*