Specificity and Promiscuity in Human Glutaminase Interacting Protein Recognition: Insight from the Binding of the Internal and C-Terminal Motif

Banerjee, Monimoy; Zoetewey, David; Ovee, Mohiuddin; Mazumder, Suman; Petrenko, Valery A.; Samoylova, Tatiana I.; Mohanty, Smita

doi:10.1021/bi3008033

Cited by 5 publications

(24 citation statements)

References 43 publications

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“…Furthermore, both DTX1 and STAU1 contain an internal motif that was previously identified as an hGIP recognition motif in a phage display library screen [6]. The demonstration of in vivo association of hGIP with partner proteins containing the same motif validates the previous discovery of S/T-X-V/L-D as a consensus internal motif for GIP recognition.…”

Section: Resultssupporting

confidence: 75%

“…However, the new partner proteins did contain an internal sequence (Fig. 1B) that matched the S/T-X-V/L-D consensus of the internal motif we had previously identified through phage display library screening [6]. …”

Section: Resultsmentioning

confidence: 92%

“…To determine whether the potential GIP-interacting partner proteins identified in the Y2H screen are recognized through classical C-terminal motif or via the recently identified novel internal GIP-recognition motif [6], sequence analyses were carried out and confirmed that the cDNAs for DTX1 and STAU1 inserted in the respective pGADT7-Rec AD library plasmids were full-length. Western blotting analysis of protein extracted from yeast transformed with the pGADT7-Rec AD library plasmids showed Gal4 AD -fusion proteins with mobilities consistent with those expected for expression of the full-length interactor proteins indicating that the authentic C-terminus of each was likely to be present on the respective fusion proteins (data not shown).…”

Section: Resultsmentioning

confidence: 99%

“…Since GIP is involved in various cancerous pathways and consists solely of a single PDZ domain, it has received considerable attention as a possible target for anticancer therapeutics [20–21]. We have been using various techniques including phage display library screening [6] and yeast two-hybrid (Y2H) assays [14] to find novel GIP interactors. Utilizing these techniques, we recently discovered that hGIP not only interacted via recognition of a C-terminal motif, but also recognized an internal motif with the consensus S/T-x-V/L-D sequence [6].…”

Section: Introductionmentioning

confidence: 99%

“…We have been using various techniques including phage display library screening [6] and yeast two-hybrid (Y2H) assays [14] to find novel GIP interactors. Utilizing these techniques, we recently discovered that hGIP not only interacted via recognition of a C-terminal motif, but also recognized an internal motif with the consensus S/T-x-V/L-D sequence [6]. …”

Section: Introductionmentioning

confidence: 99%

See 4 more Smart Citations

New partner proteins containing novel internal recognition motif for human glutaminase interacting protein (hGIP)

Zencir

Banerjee

Dobson

et al. 2013

Biochemical and Biophysical Research Communications

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Regulation of gene expression in cells is mediated by protein-protein, DNA-protein and receptor-ligand interactions. PDZ (PSD-95/Discs-large/ZO-1) domains are protein–protein interaction modules. PDZ-containing proteins function in the organization of multi-protein complexes controlling spatial and temporal fidelity of intracellular signaling pathways. In general, PDZ proteins possess multiple domains facilitating distinct interactions. The human Glutaminase Interacting Protein (hGIP) is an unusual PDZ protein comprising entirely of a single PDZ domain and plays pivotal roles in many cellular processes through its interaction with the C-terminus of partner proteins. Here, we report the identification by yeast two-hybrid screening of two new hGIP-interacting partners, DTX1 and STAU1. Both proteins lack the typical C-terminal PDZ recognition motif but contain a novel internal hGIP recognition motif recently identified in a phage display library screen. Fluorescence resonance energy transfer and confocal microscopy analysis confirmed the in vivo association of hGIP with DTX1 and STAU1 in mammalian cells validating the previous discovery of S/T-X-V/L-D as a consensus internal motif for hGIP recognition. Similar to hGIP, DTX1 and STAU1 have been implicated in neuronal function. Identification of these new interacting partners furthers our understanding of GIP-regulated signaling cascades and these interactions may represent potential new drug targets in humans.

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Section: Resultssupporting

confidence: 75%

Section: Resultsmentioning

confidence: 92%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

New partner proteins containing novel internal recognition motif for human glutaminase interacting protein (hGIP)

Zencir

Banerjee

Dobson

et al. 2013

Biochemical and Biophysical Research Communications

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Structural basis of internal peptide recognition by PDZ domains using molecular dynamics simulations

Sain

Mohanty

2019

Preprint

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PDZ domains are important peptide recognition modules which usually recognize short C-terminal stretches of their interaction partners, but certain PDZ domains can also recognize internal peptides in the interacting proteins. Due to the scarcity of data on internal peptide recognition and lack of understanding of the mechanistic details of internal peptide recognition, identification of PDZ domains capable of recognizing internal peptides has been a difficult task.Since Par-6 PDZ domain can recognize both C-terminal and internal peptides, we have carried out multiple explicit solvent MD simulations of 1 μs duration on free and peptide bound Par-6 PDZ to decipher mechanistic details of internal peptide recognition. These simulations have been analyzed to identify residues which play a crucial role in internal peptide recognition by PDZ domains. Based on the conservation profile of the identified residues, we have predicted 47 human PDZ domains to be capable of recognizing internal peptides in human. We have also investigated how binding of CDC42 to the CRIB domain adjacent to the Par6 PDZ allosterically modulate the peptide recognition by Par6 PDZ. Our MD simulations on CRIB-Par6_PDZ didomain in isolation as well as in complex with CDC42, indicate that in absence of CDC42 the adjacent CRIB domain induces open loop conformation of PDZ facilitating internal peptide recognition. On the other hand, upon binding of CDC42 to the CRIB domain, Par6 PDZ adopts closed loop conformation required for recognition of C-terminus peptides. These results provide atomistic details of how binding of interaction partners onto adjacent domains can allosterically regulate substrate binding to PDZ domains. In summary, MD simulations provide novel insights into the modulation of substrate recognition preference of PDZ by specific peptides, adjacent domains and binding of interaction partners at allosteric sites.

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Effect of Toxic Metal Binding on Tax-Interacting Protein1 (TIP1): A Protein Related to Brain Diseases

Chaudhary

Dahal

Sayania

et al. 2019

Natural Product Communications

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Human tax-interacting protein1 (TIP1), also known as glutaminase-interacting protein (GIP), is a small globular protein containing a PDZ domain. PDZ domains are the most common protein-protein interaction modules present in eukaryotes. In humans, TIP1 plays a very important role in many cellular pathways including β-catenin-mediated Wnt signaling, Rhoactivator rhotekin-mediated Rho signaling pathway, and glutamate signaling pathway for the normal activity of the central nervous system. TIP1 also regulates potassium channel expression in the plasma membrane and is a binding partner to many proteins including viral oncoproteins, HTLV-1 Tax and HPV16 E6. Since TIP1 is at a pivotal point in many cellular processes through its interaction with a growing list of partner proteins, any impact on the proper functioning of this protein can have severe consequences on the well-being of a living system. Although metals are essential for plants and animals in trace amounts, elevated levels of heavy metals such as arsenic, cadmium, zinc, and lead are toxic causing various health problems including cardiovascular disorders, neuronal damage, renal injuries, and cancer. Here, we report the effect of heavy metals, arsenic and cadmium, on TIP1 conformation using circular dichroism and fluorescence spectroscopy techniques. Our study revealed these metals have a significant impact on the structure of TIP1 even at very low levels.

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Specificity and Promiscuity in Human Glutaminase Interacting Protein Recognition: Insight from the Binding of the Internal and C-Terminal Motif

Cited by 5 publications

References 43 publications

New partner proteins containing novel internal recognition motif for human glutaminase interacting protein (hGIP)

New partner proteins containing novel internal recognition motif for human glutaminase interacting protein (hGIP)

Structural basis of internal peptide recognition by PDZ domains using molecular dynamics simulations

Effect of Toxic Metal Binding on Tax-Interacting Protein1 (TIP1): A Protein Related to Brain Diseases

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