title: An intersubunit lock-and-key 'clasp' motif in the dimer interface of Delta class glutathione transferase.
abstract: Structural investigations of a GST (glutathione transferase), adGSTD4-4, from the malaria vector Anopheles dirus show a novel lock-and-key 'Clasp' motif in the dimer interface of the Delta class enzyme. This motif also appears to be highly conserved across several insect GST classes, but differs from a previously reported mammalian lock-and-key motif. The aromatic 'key' residue not only inserts into a hydrophobic pocket, the 'lock', of the neighbouring subunit, but also acts as part of the 'lock' for the other subunit 'key'. The 'key' residues from both subunits show aromatic ring stacking with each other in a pi-pi interaction, generating a 'Clasp' in the middle of the subunit interface. Enzyme catalytic and structural characterizations revealed that single amino acid replacements in this 'Clasp' motif impacted on catalytic efficiencies, substrate selectivity and stability. Substitutions to the 'key' residue create strong positive co-operativity for glutathione binding, with a Hill coefficient approaching 2. The lock-and-key motif in general and especially the 'Clasp' motif with the pi-pi interaction appear to play a pivotal role in subunit communication between active sites, as well as in stabilizing the quaternary structure. Evidence of allosteric effects suggests an important role for this particular intersubunit architecture in regulating catalytic activity through conformational transitions of subunits. The observation of co-operativity in the mutants also implies that glutathione ligand binding and dimerization are linked. Quaternary structural changes of all mutants suggest that subunit assembly or dimerization basically manipulates subunit communication.
authors.affiliation: Institute of Molecular Biology and Genetics, Mahidol University, Salaya Campus, 25/25 Phutthamonthol Road 4, Salaya, Nakhon Pathom 73170, Thailand.
authors.forename: Jantana
authors.initials: J
authors.lastname: Wongsantichon
authors.forename: Albert J
authors.initials: AJ
authors.lastname: Ketterman
mesh_terms: D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D000852:Anopheles / Q000201:enzymology*; D001665:Binding Sites; D015971:Gene Expression Regulation, Enzymologic; D005982:Glutathione Transferase / Q000737:chemistry* / Q000145:classification* / Q000378:metabolism; D007700:Kinetics; D008958:Models, Molecular; D011485:Protein Binding; D015202:Protein Engineering; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity
chemical_list: D005982:Glutathione Transferase