Nucleotide dependent cysteine reactivity of hGBP1 uncovers a domain movement during GTP hydrolysis.
As a member of the dynamin superfamily human guanylate-binding protein 1 (hGBP1) binds and hydrolyses GTP thereby undergoing structural changes which lead to self-assembly of the protein. Here, we employ the reactivity of hGBP1 with a cysteine reactive compound in order to monitor structural changes imposed by GTP binding and hydrolysis. Positions of cysteine residues buried between the C-terminal domain of hGBP1 and the rest of the protein are identified which report a large change of accessibility by the compound after addition of GTP. Our results indicate that nucleotide hydrolysis induces a domain movement in hGBP1, which we suggest enables further assembly of the protein.
Vöpel
Tobias
T
Physikalische Chemie I, Fakultät für Chemie und Biochemie, Ruhr-Universität Bochum, 44780 Bochum, Germany.
Kunzelmann
Simone
S
Herrmann
Christian
C
D019943:Amino Acid Substitution; D001665:Binding Sites; D003545:Cysteine / Q000737:chemistry; D004228:Dithionitrobenzoic Acid; D019204:GTP-Binding Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D006160:Guanosine Triphosphate / Q000378:metabolism*; D006801:Humans; D066298:In Vitro Techniques; D007700:Kinetics; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013439:Sulfhydryl Reagents
C490287:GBP1 protein, human; D011994:Recombinant Proteins; D013439:Sulfhydryl Reagents; D006160:Guanosine Triphosphate; D004228:Dithionitrobenzoic Acid; D019204:GTP-Binding Proteins; D003545:Cysteine