Structural determinants of Cys2His2 zinc fingers.
Two mutants of the zinc finger peptide Xfin-31 (Ac-YKCGLCERSFVEKSALSRHQRVHKN-CONH2) containing alterations to the conserved hydrophobic core have been constructed and their zinc-bound structures investigated by 1H NMR techniques. In the first (Xfin-31B) a double mutation R8F/F10G places the conserved core aromatic residue at position 8 rather than position 10. In the second (Xfin-31C), Phe-10 is replaced by Leu. A qualitative analysis of 1H chemical shifts, NOE connectivities and coupling constants indicates that the global folds of both mutants are similar to that of the wild-type protein. However, amide exchange rates suggest that the F10L mutant is much less stable than either the wild-type or the R8F/F10G mutant.
Mortishire-Smith
R J
RJ
Scripps Research Institute, Department of Molecular Biology, La Jolla, CA 92037.
Lee
M S
MS
Bolinger
L
L
Wright
P E
PE
D000595:Amino Acid Sequence; D003545:Cysteine / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000235:genetics; D006639:Histidine / Q000235:genetics*; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D009154:Mutation; D010455:Peptides; D011487:Protein Conformation; D011994:Recombinant Proteins; D016335:Zinc Fingers / Q000235:genetics*
D004268:DNA-Binding Proteins; D010455:Peptides; D011994:Recombinant Proteins; C072294:zinc finger peptide Xfin-31; D006639:Histidine; D003545:Cysteine