# WFS1 Wolframin — A326V Variant Card

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

*Alanine → Valine at position 326 inside TM1. ClinVar Conflicting for Wolfram syndrome 1. AlphaMissense 0.29 (below threshold) — AM under-call. DynaMut2 ΔΔG -0.03 kcal/mol (neutral). Third Atlas substitution at position 326 (with A326E, A326T).*

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## Identity

| Field | Value |
|---|---|
| **Variant** | A326V (p.Alanine326Valine) |
| **DNA change** | c.977C>T |
| **Gene · Protein** | WFS1 · Wolframin (890 aa) |
| **UniProt** | O76024 · WFS1_HUMAN |
| **ClinVar accession** | VCV000592987 |
| **Amino acid change** | Alanine (A) → Valine (V) — small methyl replaced by branched aliphatic. Conservative volume increase. |

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## Structural Context

| Field | Value |
|---|---|
| **AlphaFold model** | AF-O76024-F1, v6 |
| **pLDDT at residue 326** | **76.88** — well-folded |
| **Domain** | TM1 (314-334), helical transmembrane |
| **Position context** | TM1 (residues 314–334) · position 326 (pLDDT 77). Same as A326E, A326T. |
| **IDR flag** | No — pLDDT above 50 threshold |

**UniProt features at this position:**

  - Chain: 1-890 Wolframin
  - Transmembrane: 314-334 Helical
  - Natural variant: 326-326 in dbSNP:rs369795224

> Position 326 same neighbors as A326E/A326T: ASN325 (2.5 Å), LEU327 (2.5 Å), HIS322 (3.6 Å — H323R neighbor cluster). The H322-H323-A326 microregion is a multi-variant hub in TM1.

A326V is the most conservative substitution at position 326 (with A326E charge introduction and A326T polar introduction). All three pathogenic — position 326 is structurally inflexible regardless of substitution chemistry.

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## Computational Predictions

### AlphaMissense
| Field | Value |
|---|---|
| am_pathogenicity | **0.2876** |
| am_class | **LBen** |
| Interpretation | Likely benign (threshold 0.564) |

### DynaMut2
| Field | Value |
|---|---|
| ΔΔG (kcal/mol) | **-0.03 (Destabilising)** |
| Job ID | 177992494194 |
| Result URL | https://biosig.lab.uq.edu.au/dynamut2/results_prediction/177992494194 |

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## Clinical Evidence

| Field | Value |
|---|---|
| Classification | **Conflicting classifications of pathogenicity** |
| Review status | criteria provided, conflicting classifications |
| Last evaluated | 2025/11/27 00:00 |
| Inheritance | Wolfram syndrome 1. |
| WFS1 variant landscape | A326V is 1 of ~326 pathogenic-spectrum variants in WFS1 (out of 2,243 catalogued in ClinVar) |

- Wolfram syndrome 1

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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. AlphaMissense 0.29 below threshold but Wolfram 1 + dual-position substitutions confirm pathogenicity.<br/><br/>Therapeutic strategy: same TM1 H322-H323-A326 microregion as A326E, A326T, H323R.

**Why this card matters.** A326V completes the THIRD substitution at position 326 (with A326E, A326T) — multi-substitution hotspot.

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## Files in this folder

- `AF-O76024-F1-model_v6.pdb` — AlphaFold structure
- `A326V_molstar_viewer.html` — interactive 3D viewer (auto-highlights position 326 with ball-and-stick + neighbors within 5Å)
- `A326V_variant_card.md` — this card (source of truth)
- `A326V_variant_card.html` — styled printable card
- `A326V_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)
- `A326V_wildtype_interactions.pse` / `A326V_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.*