title: Specificity and promiscuity in human glutaminase interacting protein recognition: insight from the binding of the internal and C-terminal motif.
abstract: A large number of cellular processes are mediated by protein-protein interactions, often specified by particular protein binding modules. PDZ domains make up an important class of protein-protein interaction modules that typically bind to the C-terminus of target proteins. These domains act as a scaffold where signaling molecules are linked to a multiprotein complex. Human glutaminase interacting protein (GIP), also known as tax interacting protein 1, is unique among PDZ domain-containing proteins because it is composed almost exclusively of a single PDZ domain rather than one of many domains as part of a larger protein. GIP plays pivotal roles in cellular signaling, protein scaffolding, and cancer pathways via its interaction with the C-terminus of a growing list of partner proteins. We have identified novel internal motifs that are recognized by GIP through combinatorial phage library screening. Leu and Asp residues in the consensus sequence were identified to be critical for binding to GIP through site-directed mutagenesis studies. Structure-based models of GIP bound to two different surrogate peptides determined from nuclear magnetic resonance constraints revealed that the binding pocket is flexible enough to accommodate either the smaller carboxylate (COO(-)) group of a C-terminal recognition motif or the bulkier aspartate side chain (CH(2)COO(-)) of an internal motif. The noncanonical ILGF loop in GIP moves in for the C-terminal motif but moves out for the internal recognition motifs, allowing binding to different partner proteins. One of the peptides colocalizes with GIP within human glioma cells, indicating that GIP might be a potential target for anticancer therapeutics.
authors.affiliation: Department of Chemistry and Biochemistry, Auburn University, Auburn, AL 36849, USA.
authors.forename: Monimoy
authors.initials: M
authors.lastname: Banerjee
authors.forename: David L
authors.initials: DL
authors.lastname: Zoetewey
authors.forename: Mohiuddin
authors.initials: M
authors.lastname: Ovee
authors.forename: Suman
authors.initials: S
authors.lastname: Mazumder
authors.forename: Valery A
authors.initials: VA
authors.lastname: Petrenko
authors.forename: Tatiana I
authors.initials: TI
authors.lastname: Samoylova
authors.forename: Smita
authors.initials: S
authors.lastname: Mohanty
mesh_terms: D000595:Amino Acid Sequence; D045744:Cell Line, Tumor; D005910:Glioma / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000032:analysis / Q000737:chemistry / Q000378:metabolism*; D008958:Models, Molecular; D054731:PDZ Domains; D019151:Peptide Library; D010455:Peptides / Q000032:analysis / Q000737:chemistry* / Q000378:metabolism*; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs
chemical_list: D047908:Intracellular Signaling Peptides and Proteins; D019151:Peptide Library; D010455:Peptides; C421249:TAX1BP3 protein, human