Skip to main content

Table 1 List of the target triplets, amino acid residues at the position −1 ~ +6 of the α-helix of each single-zinc-finger domain of active 7ZFPs and their binding activity

From: C-terminal in Sp1-like artificial zinc-finger proteins plays crucial roles in determining their DNA binding affinity

Active 7ZFPs and their variants marked with the line number p16 DNA binding activitya N-terminal Sp1three-zinc-finger domain (3ZF) and its mutants Middle Sp1-liketwo-zinc-finger modules (2ZF) C-terminal Sp1-liketwo-zinc-finger modules (2ZF) C-terminal linkerbetween finger-7& 6 × His-tag
finger-1 finger-2 finger-3 finger-4 finger-5 finger-6 finger-7
5′-ggg-3′ 5′-gcg-3′ 5′-ggg-3′ 5′-aac-3′ 5′-gga-3′ 5′-gaa-3′ 5′-gag-3′
Finger motif replacement
1_ Active-A (consensus)b (++) KTSHLRA RSDELQR RSDHLSK ESDNLSQ QSGHLQR QSSNLQR RSDNLAR TGEKP
2_Variant-A (f3) (++) −−−−−−− c −−−−−−− Q−G−−QR −−−−−−− −−−−−−− −−−−−−− −−−−−−− −−−−−
3_ Active-B (++) −−−−−−− −−−−−−− −−−−−−− −−−−−−− −−−−−−− −−−−−−− −−−−−T− −−−−−
4_Variant-B (f3) (++) −−−−−−− −−−−−−− Q−G−−QR −−−−−−− −−−−−−− −−−−−−− −−−−−T− −−−−−
5_Variant-B (f6f7) (−) −−−−−−− −−−−−−− −−−−−−− −−−−−−− −−−−−−− D−G−−R− Q−GH−Q− −−−−−
6_ Active-C (++) −−−−−−− −−−−−−− −−−−−−− −−−−−−− −−T−−−− −−−−−−− −−−−−T− −−−−−
7_Variant-C (f6f7) (−) −−−−−−− −−−−−−− −−−−−−− −−−−−−− −−T−−−− D−G−−R− Q−GH−Q− −−−−−
Single key amino acid mutation
8_ Active-D (+) −−−−−−− −−−−−−− −−−−−−− −−−−−−− −−−−−−− −−−−−V− −−−A−Q− −−−−−
9_Variant-D (f6) (−) −−−−−−− −−−−−−− −−−−−−− −−−−−−− −−−−−−− −−R−−V− −−−A−Q− −−−−−
10_ Active-F (++) −−−−−−− −−−−−−− −−−−−−− −−−−−−− −−T−−−− −−−−−V− −−−−−T− −−−−−
11_Variant-F (f6)d (−) −−−−−−− −−−−−−− −−−−−−− −−−−−−− −−T−−−− −−R−−V − −−−−−T− −−−−−
12_ Active-G (++) −−−−−−− −−−−−−− −−−−−−− D−G−−RV −−−−−−− −−−−−V− −−−−−T− −−−−−
13_Variant-G (f6) (−) −−−−−−− −−−−−−− −−−−−−− D−G−−RV −−−−−−− −−R−−V − −−−−−T− −−−−−
14_ Active-Hc (+) −−−−−−− −−−−−−− −−−−−−− D−G−−RV −−−−−−− −−−−−V− −−−A−Q− −−−−−
15_Variant-H (f6) (−) −−−−−−− −−−−−−− −−−−−−− D−G−−RV −−−−−−− −−R−−V− −−−A−Q− −−−−−
16_ Active-I (++) −−−−−−− −−−−−−− −−−−−−− D−G−−RR −−−−−−− −−−−−V− −−−−−T− −−−−−
17_Variant-I (f6) (−) −−−−−−− −−−−−−− −−−−−−− D−G−−RR −−−−−−− −−R−−V − −−−−−T− −−−−−
18_ Active-J (+) −−−−−−− −−−−−−− −−−−−−− D−G−−RR −−T−−−− −−−−−V− −−−A−Q− −−−−−
19_Variant-J (f6) (−) −−−−−−− −−−−−−− −−−−−−− D−G−−RR −−T−−−− −−R−−V− −−−A−Q− −−−−−
20_ Active-K (++) −−−−−−− −−−−−−− −−−−−−− D−G−−RR −−T−−−− −−−−−V− −−−−−T− −−−−−
21_Variant-K (f6) (−) −−−−−−− −−−−−−− −−−−−−− D−G−−RR −−T−−−− −−R−−V − −−−−−T− −−−−−
C -terminal linker deletion
22_ Active-K (++) −−−−−−− −−−−−−− −−−−−−− −−−−−−− −−−−−−− −−−−−V− −−−−−T− −−−−−
23_Variant-K (linker) (−) −−−−−−− −−−−−−− −−−−−−− −−−−−−− −−−−−−− −−−−−V− −−−−−T− TG
  1. aanalyzed in EMSA assays.
  2. bthe exact sequence of amino acids of 7ZFP Active-A: N-MASRDPGKKKQHICHIQGCGKVYGKTSHLRAHLRWHTGERPFMCTWSYCGKRFTRSDELQRHKRTHTGEKKFACPECPKRFMRSDHLSKHIKTHQTGEKPFMCTWSYCGKRFTESDNLSQHKRTHTGEKKFACPECPKRFMQSGHLQRHIKTHTGEKPFMCTWSYCGKRFTQSSNLQRHKRTHTGEKKFACPECPKRFMRSDNLARHIKTHTGEKPFLEHHHHHH-C.
  3. cthe symbol “−” means the amino acid residue is the same as that in the consensus sequence of Active-A at the same position in the same column. dused in optimization assays for soluble 7ZFP preparation.