Oxidation of a critical methionine modulates DNA binding of the Drosophila melanogaster high mobility group protein, HMG-D.
HMG-D is a major high mobility group chromosomal protein present during early embryogenesis in Drosophila melanogaster. During overexpression and purification of HMG-D from E. coli, a key DNA binding residue, methionine 13, undergoes oxidation to methionine sulfoxide. Oxidation of this critical residue decreases the affinity of HMG-D for DNA by three-fold, altering the structure of the HMG-D-DNA complex without affecting the structure of the free protein. This work shows that minor modification of DNA intercalating residues may be used to fine tune the DNA binding affinity of HMG domain proteins.
Dow
L K
LK
Department of Cell and Structural Biology, University of Illinois at Urbana-Champaign, Urbana 61801, USA.
Changela
A
A
Hefner
H E
HE
Churchill
M E
ME
D000818:Animals; D001665:Binding Sites; D004247:DNA / Q000378:metabolism*; D004220:Disulfides; D004331:Drosophila melanogaster / Q000737:chemistry*; D004586:Electrophoresis / Q000379:methods; D004926:Escherichia coli / Q000235:genetics; D006609:High Mobility Group Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy; D013058:Mass Spectrometry; D008715:Methionine / Q000378:metabolism; D009692:Nucleic Acid Heteroduplexes; D010084:Oxidation-Reduction; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
D004220:Disulfides; D006609:High Mobility Group Proteins; D009692:Nucleic Acid Heteroduplexes; D011994:Recombinant Proteins; C104923:high mobility group protein D; D004247:DNA; D008715:Methionine