Table 4. Summary of protein stability and functions of UBA5 variants.

	Variants	Thermal stability	ATP-binding defect ‡	UFM1 activation	UFM1 transthiolation
Group IA	p.Ala371Thr	Decreased	No defect	Normal	Decreased
	p.Asp389Gly	Normal	No defect	Decreased	Decreased
	p.Asp389Tyr	Normal	No defect	Decreased	Normal
Group IB	p.Arg72Cys	Decreased	No defect	Decreased	Normal
	p.Gln312Leu	Local destabilization†	No defect	Normal	Normal
Group II	p.Tyr53Phe	Normal	Strong	Decreased	N.A.
	p.Met57Val	Normal	Intermediate	Normal	Normal
	p.Val260Met	Decreased	Intermediate	Decreased	N.A.
Group III	p.Arg55His	Normal	Strong	Near baseline	N.A.
	p.Gly168Glu	N.A.*	N.A.	N.A.	N.A.
	p.Leu254Pro	Decreased	Strong	Near baseline	N.A.
	p.Cys303Arg	N.A.*	N.A.	N.A.	N.A.

^*Insoluble in protein purification.

^†See Results and Figure 6B.

^‡Strong, 4–5°C in Tm shift; intermediate, 6–8°C in Tm shift. Reference UBA5 shows 13 °C in Tm shift upon ATP addition.