Table 2.

Protein stability analysis of MKRN3 with missense variants

Mutation (missense)	ΔΔG	Protein stability	Location within the protein
p.Arg68His	0.915	Slightly destabilizing	Before C3H1 zinc finger 1
p.Arg242Trp	–1.090	Stabilizing	Second C3H1 zinc finger
p.Met297Arg	–0.240	Neutral	Between MRKN type Cys-His domain and C3HC4 ring finger
p.Ala302Val	1.000	Destabilizing	Between MRKN type Cys-His domain and C3HC4 ring finger
p.Arg328His	1.409	Destabilizing	C3HC4 ring finger
p.Arg328Cys	1.661	Destabilizing	C3HC4 ring finger
p.Arg365Ser	1.617	Destabilizing	C3HC4 ring finger
p.Phe417Ile	–1.010	Stabilizing	Third C3H1 zinc finger
p.Phe417Leu	–1.392	Stabilizing	Third C3H1 zinc finger
p.Tyr418His	2.882	Highly destabilizing	Third C3H1 zinc finger

ΔΔG is the energy difference between the wild type and a variant of MKRN3. This analysis can provide a classification of the mutation into 7 categories: (1) highly stabilizing (ΔΔG < −1.84 kcal/mol); (2) stabilizing (−1.84 kcal/mol ≤ ΔΔG < −0.92 kcal/mol); (3) slightly stabilizing (−0.92 kcal/mol ≤ ΔΔG < −0.46 kcal/mol); (4) neutral (−0.46 kcal/mol < ΔΔG ≤ +0.46 kcal/mol); (5) slightly destabilizing (+0.46 kcal/mol < ΔΔG ≤ +0.92 kcal/mol); (6) destabilizing (+0.92 kcal/mol < ΔΔG ≤ +1.84 kcal/mol); and (7) highly destabilizing (ΔΔG > +1.84 kcal/mol) (11).