Table 2.

Amino acid composition of NtAQPs at known functionally important positions

	Gene Name	NPA I	NPA II	ar/R					Froger's positions					Other regulatory residues							Pred. subcellular loc.
		LB	LE	H2	LC	H5	LE1	LE2	P1	P2	P3	P4	P5	Asp28	Glu31	Ser115	Ser188	His193	Ser274	Ser277	mPLoc	WolfPsort	Yloc
PIP	NtPIP1;1s	NPA	NPA	F	G	H	T	R	M	S	A	F	W	E	E	SP	N	H	-	S	plas	plas	cyto
	NtPIP1;1t	NPA	NPA	F	G	H	T	R	M	S	A	F	W	E	E	SP	N	H	-	S	plas	plas	cyto
	NtPIP1;2s	NPA	NPA	F	G	H	T	R	Q	S	A	F	W	E	E	SP	N	H	-	S	plas	plas	cyto
	NtPIP1;2t	NPA	NPA	F	G	H	T	R	Q	S	A	F	W	E	E	SP	N	H	-	S	plas	plas	cyto
	NtPIP1;3s	NPA	NPA	F	G	H	T	R	M	S	A	F	W	E	E	SP	N	H	-	S	plas	plas	cyto
	NtPIP1;3t	NPA	NPA	F	G	H	T	R	M	S	A	F	W	E	E	SP	N	H	-	S	plas	plas	cyto
	NtPIP1;5s	NPA	NPA	F	G	H	T	R	M	S	A	F	W	E	E	SP	N	H	-	S	plas	plas	cyto
	NtPIP1;5t	NPA	NPA	F	G	H	T	R	M	S	A	F	W	E	E	SP	N	H	-	S	plas	plas	cyto
	NtPIP1;7t	NPA	NPA	F	G	H	T	R	G	S	A	F	W	E	E	SP	SP	H	-	S	plas	plas	cyto
	NtPIP1;8s	NPA	NPA	F	G	H	T	R	G	S	A	F	W	E	E	SP	SP	H	-	S	plas	plas	plas
	NtPIP2;1s	NPA	NPA	F	G	H	T	R	Q	S	A	F	W	D	E	SP	N	H	SP	S	plas	plas	plas
	NtPIP2;1x	NPA	NPA	F	G	H	T	R	Q	S	A	F	W	D	E	SP	N	H	SP	S	plas	plas	plas
	NtPIP2;2t	NPA	NPA	F	G	H	T	R	Q	S	A	F	W	D	E	SP	N	H	SP	S	plas	plas	plas
	NtPIP2;3t	NPA	NPA	F	G	H	T	R	Q	S	A	F	W	D	E	SP	N	H	SP	S	plas	plas	plas
	NtPIP2;4s	NPA	NPA	F	G	H	T	R	Q	S	A	F	W	D	E	SP	N	H	SP	S	plas	plas	plas
	NtPIP2;4t	NPA	NPA	F	G	H	T	R	Q	S	A	F	W	D	E	SP	N	H	SP	S	plas	plas	plas
	NtPIP2;5s	NPA	NPA	F	G	H	T	R	Q	S	A	F	W	D	E	SP	N	H	SP	S	plas	plas	plas
	NtPIP2;5t	NPA	NPA	F	G	H	T	R	Q	S	A	F	W	D	E	SP	N	H	SP	S	plas	plas	plas
	NtPIP2;6s	NPA	NPA	F	G	H	T	R	Q	S	A	F	W	D	E	SP	SP	H	SP	S	plas	plas	plas
	NtPIP2;6t	NPA	NPA	F	G	H	T	R	Q	S	A	F	W	D	E	SP	SP	H	SP	S	plas	plas	plas
	NtPIP2;7t	NPA	NPA	F	G	H	T	R	Q	S	A	F	W	D	E	SP	SP	H	SP	S	plas	plas	plas
	NtPIP2;8s	NPA	NPA	F	G	H	T	R	M	S	A	F	W	D	E	S	SP	H	SP	S	plas	plas	plas
	NtPIP2;8t	NPA	NPA	F	G	H	T	R	M	S	A	F	W	D	E	S	SP	H	SP	S	plas	plas	plas
	NtPIP2;9s	NPA	NPA	F	G	H	T	R	M	S	A	F	W	D	E	SP	SP	H	SP	S	plas	plas	plas
	NtPIP2;9t	NPA	NPA	F	G	H	T	R	M	S	A	F	W	D	E	SP	SP	H	SP	S	plas	plas	plas
	NtPIP2;11s	NPA	NPA	F	G	H	T	R	M	S	A	F	W	D	E	SP	N	H	SP	S	plas	plas	plas
	NtPIP2;11t	NPA	NPA	F	G	H	T	R	M	S	A	F	W	D	E	SP	N	H	SP	S	plas	plas	plas
	NtPIP2;13s	NPA	NPA	F	G	H	T	R	Q	S	A	F	W	D	E	SP	N	H	SP	SP	plas	plas	plas
	NtPIP2;13t	NPA	NPA	F	G	H	T	R	Q	S	A	F	W	D	E	SP	N	H	SP	SP	plas	plas	plas
NIP	NtNIP1;1s	NPS	NPA	W	A	V	A	R	F	S	A	Y	L	-	-	P	-	-	SP	-	plas	plas	cyto
	NtNIP1;2s	NPA	NPA	W	L	V	A	R	F	S	A	Y	M	-	-	P	-	-	S	-	plas	plas	plas
	NtNIP1;2t	NPA	NPA	W	L	V	A	R	F	S	A	Y	M	-	-	P	-	-	S	-	plas	plas	plas
	NtNIP2;1s	NPA	NPA	G	T	S	G	R	L	T	A	Y	I	-	-	P	-	-	SP	-	plas	plas	plas
	NtNIP3;1s	NPA	NPA	W	V	I	A	R	F	S	A	Y	I	-	-	P	-	-	S	-	plas	plas	plas
	NtNIP4;1s	NPA	NPA	W	F	V	A	R	F	S	A	Y	I	-	-	P	-	-	S	-	plas	plas	plas
	NtNIP4;1t	NPA	NPA	W	F	V	A	R	F	S	A	Y	I	-	-	P	-	-	S	-	plas	plas	plas
	NtNIP4;2s	NPA	NPA	W	F	V	A	R	L	S	A	Y	I	-	-	P	-	-	T	-	plas	plas	plas
	NtNIP4;2t	NPA	NPA	W	F	V	A	R	L	S	A	Y	I	-	-	P	-	-	T	-	plas	plas	plas
	NtNIP4;3s	NPA	NPA	W	F	V	A	R	L	S	A	Y	I	-	-	SP	-	-	S	-	plas	tono	plas
	NtNIP5;1s	NPA	NPA	S	V	I	A	R	F	T	A	Y	I	-	-	P	-	-	S	-	plas	tono	cyto
	NtNIP5;1t	NPS	NPV	A	V	I	A	R	F	T	A	Y	L	-	-	P	-	-	S	-	plas	tono	cyto
	NtNIP6;1s	NPA	NPV	S	V	I	A	R	L	T	A	Y	L	-	-	P	-	-	S	-	plas	plas	plas
	NtNIP6;1t	NPA	NPV	T	V	I	A	R	L	T	A	Y	L	-	-	P	-	-	S	-	plas	plas	plas
	NtNIP7;1s	NPA	NPA	A	I	V	G	R	Y	S	A	Y	V	-	-	P	-	-	T	-	plas	plas	plas
	NtNIP7;1t	NPA	NPA	A	I	V	G	R	Y	S	A	Y	V	-	-	P	-	-	T	-	plas	plas	plas
TIP	NtTIP1;1s	NPA	NPA	H	F	I	A	V	A	S	A	Y	W	-	-	T	-	-	-	-	tono	plas	plas
	NtTIP1;1t	NPA	NPA	H	F	I	A	V	A	S	A	Y	W	-	-	T	-	-	-	-	tono	plas	plas
	NtTIP1;2s	NPA	NPA	H	F	I	A	V	A	S	A	Y	W	-	-	T	-	-	-	-	tono	plas	pero
	NtTIP1;2t	NPA	NPA	H	F	I	A	V	A	S	A	Y	W	-	-	T	-	-	-	-	tono	plas	pero
	NtTIP1;3s	NPA	NPA	H	F	I	A	V	T	S	A	Y	W	-	-	T	-	-	-	-	tono	plas	plas
	NtTIP1;3t	NPA	NPA	H	F	I	A	V	T	S	A	Y	W	-	-	T	-	-	-	-	tono	plas	plas
	NtTIP1;4t	NPA	NPA	H	F	I	A	V	A	S	A	Y	W	-	-	T	-	-	-	-	tono	plas	cyto
	NtTIP2;1s	NPD	NPA	H	H	I	G	R	V	S	A	Y	W	-	-	T	-	-	-	-	tono	tono	plas
	NtTIP2;1t	NPA	NPA	H	H	I	G	R	V	S	A	Y	W	-	-	T	-	-	-	-	tono	plas	plas
	NtTIP2;2s	NPA	NPA	H	H	I	G	R	V	S	A	Y	W	-	-	T	-	-	-	-	tono	tono	plas
	NtTIP2;3s	NPA	NPA	H	H	I	G	R	V	S	A	Y	W	-	-	T	-	-	-	-	tono	tono	Extra
	NtTIP2;3t	NPA	NPA	H	H	I	G	R	V	S	A	Y	W	-	-	T	-	-	-	-	tono	tono	plas
	NtTIP2;4s	NPA	NPA	H	H	I	G	R	V	S	A	Y	W	-	-	T	-	-	-	-	tono	plas	plas
	NtTIP2;5s	NPA	NPA	H	H	I	G	R	V	S	A	Y	W	-	-	T	-	-	-	-	tono	plas	plas
	NtTIP2;5t	NPA	NPA	H	H	I	G	R	V	S	A	Y	W	-	-	T	-	-	-	-	tono	plas	plas
	NtTIP3;1s	NPA	NPA	H	F	I	A	R	A	A	A	Y	W	-	-	SP	-	-	-	-	tono	plas	plas
	NtTIP3;1t	NPA	NPA	H	F	I	A	R	A	A	A	Y	W	-	-	SP	-	-	-	-	tono	plas	plas
	NtTIP3;2t	NPA	NPA	H	F	V	G	R	A	A	A	Y	W	-	-	SP	-	-	-	-	tono	plas	plas
	NtTIP4;1s	NPA	NPA	H	H	I	A	R	V	S	A	Y	W	-	-	T	-	-	-	-	tono	tono	plas
	NtTIP4;1t	NPA	NPA	H	H	I	A	R	L	S	A	Y	W	-	-	T	-	-	-	-	tono	tono	plas
	NtTIP5;1s	NPA	NPA	N	H	V	G	Y	T	S	A	Y	W	-	-	S	-	-	-	-	tono	plas	plas
	NtTIP5;1t	NPA	NPA	N	H	V	G	Y	T	S	A	Y	W	-	-	S	-	-	-	-	plas, vac	plas	plas
SIP	NtSIP1;1t	NPT	NPA	T	E	V	P	N	M	A	A	Y	W	-	-	H	-	-	-	-	plas	plas	Extra
	NtSIP1;2s	NPA	NPA	T	G	V	P	N	I	A	A	Y	W	-	-	T	-	-	-	-	plas	tono	Extra
	NtSIP1;2t	NPA	NPA	T	G	V	P	N	I	A	A	Y	W	-	-	T	-	-	-	-	plas	tono	ER
	NtSIP2;1s	NPL	NPA	H	R	H	G	S	F	V	A	Y	W	-	-	S	-	-	-	-	plas	chlo	Extra
	NtSIP2;1t	NPL	NPA	H	R	H	G	S	F	V	A	Y	W	-	-	S	-	-	-	-	plas	chlo	Extra
XIP	NtXIP1;6t	NPV	NPA	A	-	T	A	R	V	C	A	F	W	-	-	S	-	-	-	-	plas	plas	plas
	NtXIP1;6s	NPV	NPA	A	-	T	A	R	V	C	A	F	W	-	-	S	-	-	-	-	plas	plas	plas
	NtXIP1;7s	NPV	NPA	I	-	T	A	R	V	C	A	F	W	-	-	S	-	-	-	-	plas	plas	pero
	NtXIP1;7t	NPV	NPA	I	-	T	A	R	V	C	A	F	W	-	-	S	-	-	-	-	plas	plas	pero

Amino acid composition of NtAQPs at known functionally important positions. Listed are the two NPA motifs (NPA I in Loop B, NPA II in Loop E), ar/R residues (in Helix 2, 5, Loop C and Loop E) and the Froger’s residues (Positions 1-5). Other known regulatory residues involved with post-translational regulation of AQPs are listed: Asp28, Glu31, Ser115, His131, His193, Ser274, Ser277. Amino acid positions of these sites are relative to Spinach PIP2;1 or Arabidposis TIP2;1 (LC - His131 residue), as their crystal structures have been defined and well-studied. Serine (S) residues with a superscript P (i.e. SP) are predicted to be phosphorylated according to NetPhos3.1. Also included are the predicted subcellular localisations from Plant-mPloc, WolfPsort and YLoc; outputs include plasma membrane (plas), cytosol (cyto), tonoplast (tono), chloroplast (chlo), endoplasmic reticulum (ER), peroxisome (pero) and extracellular (extra) localisation