Experiments

	Mutation name	Amino acid position	Residual activity*	Intracellular processing*	Secreted into medium*	Intracellular localization*	3D-model
-	p.Cys55Phe	55	No	No	No	ER	Disulfide on C55-C358 locks initial β-strand on B-peptide (α/β)
-	p.His72Leu	72	No	Yes	Yes	Lysosomal,Lysosomal	Active site, Zn++ coordination (α/β)
-	p.Asp74Glu	74	No	Yes	Yes	Lysosomal	Active site, Zn++ coordination (α/β)
-	p.Ala95Pro	95	No	Abnormal	Yes	ER	Loop over the active site; dimer interface (α/β)
-	p.Tyr99His	99	No	Yes	Yes	Lysosomal	Dimer interface (α/β)
-	p.Asp102Asn	102	No	Yes	Yes	Lysosomal	Folding of the α/β-domain
-	p.Gly153Val	153	No	Yes	Yes, weak	Lysosomal	Destabilisation of the α/β-domain and interface to β3
-	p.Asp159Asn	159	No	Yes	Yes	Lysosomal	Turn between barrel strand and helix (α/β)
-	p.Pro197Arg	197	No	No	Yes	ER	Active site area (α/β)
-	p.His200Leu	200	Some	Yes	Yes	Lysosomal	Active site area (α/β)
-	p.His200Asn	200	Yes	Yes	Yes	Lysosomal	Active site area (α/β)
-	p.Arg202Pro	202	Some	Yes	Yes	ER	Active site area (α/β)
-	p.Arg229Trp	229	Yes	Yes	Yes	Lysosomal	Surface helix, interior (α/β)
-	p.Pro248Leu	248	Yes	Yes,Yes	Yes	-	Exposed on surface, Tolerated
-	p.Val256_Gly260del5	256	No	No	Yes	ER	-
-	p.Pro263Leu	263	No	Yes	Yes	Lysosomal	Two adjacent prolines preceding disulfide C268-C273 (α/β)
-	p.Leu278Val	278	Yes	-	Yes	-	Buried in surface loop, Tolerated
-	p.Pro282Ser	282	Yes	Yes	Yes	-	Exposed on surface, Tolerated
-	p.Thr312Ile	312	Yes	Yes	Yes	-	Buried, Tolerated
-	p.Ser318Leu	318	Yes	Yes, weak	Yes	Lysosomal	Surface, close to disulfide C268-C273 (α/β)
-	p.Arg337Gln	337	Yes	Yes	Yes	-	Exposed on surface, Tolerated
-	p.Gln339_Val342del4	339	No	No	No	ER	-
-	p.Leu352Pro	352	No	No	No	ER	Destabilisation of β-sheet below N-terminus (α/β)
-	p.Thr355Pro	355	No	No	No	ER	Preceding α-helix and disulphide bond C55-C358 (α/β)
-	p.Pro356Arg	356	No	No	No	ER	Initiation of α-helix and disulphide bond C55-C358 (α/β)
-	p.Pro379Leu	379	No	Yes	Yes	Lysosomal	Preceding a loop to the active site area (a/ß)
-	p.Gly390Cys	390	No	No	No	ER	Interface between a/ß and ß2 domain
-	p.Glu402Lys	402	Yes	Yes	Yes	Lysosomal	Buried, Tolerated
-	p.Asn413Ser	413	Yes	Yes	Yes	-	Exposed on surface, Tolerated
-	p.Gly420Val	420	No	Abnormal	No	Lysosomal	Turn preceding the proteolytic site between B and C-peptides (3α)
-	p.His445Tyr	445	No	No	Yes	ER	Active site area (3α)
-	p.Gly451Cys	451	Yes	Yes, weak	Yes	Lysosomal	Active site area (3α)
-	p.Ser453Tyr	453	No	Yes, weak	Yes	ER	Active site area (3α)
-	p.Ser453Phe	453	No	No	Yes	ER	Active site area (3α)
-	p.Val457Glu	457	Yes	Yes, weak	Yes	Lysosomal	Active site area, right behind Trp77 (3α)
-	p.Ala481Ser	481	Yes	No	Yes	-	Exposed on surface, Tolerated
-	p.Cys501Ser	501	Yes	Abnormal	Yes	Lysosomal	Disulfide C493-C501 supports conserved glycosylation at Asn497
-	p.Leu565Pro	565	Some	No	No	ER	Destabilisation of β-sheet (β1)
-	p.Pro669Leu	669	Yes	Yes	Yes	-	Buried in surface structure, Tolerated
-	p.Trp714Arg	714	No	No	No	ER	Interior of large β-domain (β2)
-	p.Thr745Arg	745	No	Yes, weak	No	Lysosomal	Interior of large β-domain (β2)
-	p.Arg750Trp	750	No	No	No	ER (lamp),ER (pdi)	Domain interface from β2 to α/β and β3)
-	p.Gly800Trp	800	No	No	Yes	ER	β sheet double layer (β2)
-	p.Gly800Arg	800	No	No	Yes	ER	β sheet double layer (β2)
-	p.Leu809Pro	809	No	No	No	ER	Destabilisation of β-sheet (β2)
-	p.His814_Arg815dup	814	No	Yes	Yes	ER	HR duplication on loop towards α/β (β2)
-	p.Gly891Arg	891	No	No	No	ER	Hinge between β1 and β3
-	p.Leu892Pro	892	No	No	No	ER	Hinge between β1 and β3
-	p.Arg916His	916	No	No	No	ER	Domain interface between β3 and α/β (β3)
-	p.Arg916Cys	916	No	No	No	ER	Domain interface between β3 and α/β (β3)
-	p.Arg950Pro	950	No	Yes	No	Lysosomal	Surface residue, proline destabilises β-sheet (β3)
-	p.Leu956Arg	956	No	No	No	ER	Domain interface between β3 and α/β (β3)
-	p.Arg962His	962	Yes	No	Yes	Lysosomal	Interaction between β3 and the α/β-domain
-	p.Phe1000Ser	1000	No	Yes, weak	No	ER	Core of ß3

*= in transfected mammalian cells