Solution NMR characterization of Sgf73(1-104) indicates that Zn ion is required to stabilize zinc finger motif.
Zinc finger motif contains a zinc ion coordinated by several conserved amino acid residues. Yeast Sgf73 protein was identified as a component of SAGA (Spt/Ada/Gcn5 acetyltransferase) multi-subunit complex and Sgf73 protein was known to contain two zinc finger motifs. Sgf73(1-104), containing the first zinc finger motif, was necessary to modulate the deubiquitinase activity of SAGA complex. Here, Sgf73(1-104) was over-expressed using bacterial expression system and purified for solution NMR (nuclear magnetic resonance) structural studies. Secondary structure and site-specific relaxation analysis of Sgf73(1-104) were achieved after solution NMR backbone assignment. Solution NMR and circular dichroism analysis of Sgf73(1-104) after zinc ion removal using chelation reagent EDTA (ethylene-diamine-tetraacetic acid) demonstrated that zinc ion was required to maintain stable conformation of the zinc finger motif.
Lai
Chaohua
C
National Laboratory for Physical Science at Microscale, University of Science and Technology of China, Hefei, Anhui 230026, PR China.
Wu
Minhao
M
Li
Pan
P
Shi
Chaowei
C
Tian
Changlin
C
Zang
Jianye
J
D000595:Amino Acid Sequence; D002942:Circular Dichroism; D004492:Edetic Acid; D004795:Enzyme Stability; D051548:Histone Acetyltransferases / Q000737:chemistry* / Q000235:genetics; D008969:Molecular Sequence Data; D019906:Nuclear Magnetic Resonance, Biomolecular; D017433:Protein Structure, Secondary; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D012996:Solutions; D015032:Zinc / Q000737:chemistry*; D016335:Zinc Fingers
D012996:Solutions; D004492:Edetic Acid; D051548:Histone Acetyltransferases; C515833:Sgf73 protein, S cerevisiae; D015032:Zinc