Flavonoids from Radix Scutellariae as potential stroke therapeutic agents by targeting the second postsynaptic density 95 (PSD-95)/disc large/zonula occludens-1 (PDZ) domain of PSD-95

PDZ (PSD-95/Disc large/Zonula occludens-1) protein interaction domains bind to cytoplasmic protein C-termini of transmembrane proteins. In order to identify new interaction partners of the voltage-gated L-type Ca2+ channel 1.2 and the plasma membrane Ca2+ ATPase 4b (PMCA4b), we used PDZ domain arrays probing for 124 PDZ domains. We confirmed this by GST pull-downs and immunoprecipitations. In PDZ arrays, strongest interactions with 1.2 and PMCA4b were found for the PDZ domains of SAP-102, MAST-205, MAGI-1, MAGI-2, MAGI-3, and ZO-1. We observed binding of the 1.2 C-terminus to PDZ domains of NHERF1/2, Mint-2, and CASK. PMCA4b was observed to interact with Mint-2 and its known interactions with Chapsyn-110 and CASK were confirmed. Furthermore, we validated interaction of 1.2 and PMCA4b with NHERF1/2, CASK, MAST-205 and MAGI-3 via immunoprecipitation. We also verified the interaction of 1.2 and nNOS and hypothesized that nNOS overexpression might reduce Ca2+ influx through 1.2. To address this, we measured Ca2+ currents in HEK 293 cells co-expressing 1.2 and nNOS and observed reduced voltage-dependent 1.2 activation. Taken together, we conclude that 1.2 and PMCA4b bind promiscuously to various PDZ domains, and that our data provides the basis for further investigation of the physiological consequences of these interactions.

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Protein Kinase D Intracellular Localization and Activity Control Kinase D-interacting Substrate of 220-kDa Traffic through a Postsynaptic Density-95/Discs Large/Zonula Occludens-1-binding Motif

Journal of Biological Chemistry ◽

10.1074/jbc.m603044200 ◽

2006 ◽

Vol 281 (27) ◽

pp. 18888-18900 ◽

Cited By ~ 53

Author(s):

Lucía Sánchez-Ruiloba ◽

Noemí Cabrera-Poch ◽

María Rodríguez-Martínez ◽

Celia López-Menéndez ◽

Roberto Martín Jean-Mairet ◽

...

Keyword(s):

Protein Kinase ◽

Intracellular Localization ◽

Postsynaptic Density ◽

Protein Kinase D ◽

Binding Motif ◽

Zonula Occludens ◽

Discs Large ◽

Zonula Occludens 1

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Ankrd1-mediated signaling is supported by its interaction with zonula occludens-1

Archives of Biological Sciences ◽

10.2298/abs1403233n ◽

2014 ◽

Vol 66 (3) ◽

pp. 1233-1242 ◽

Cited By ~ 2

Author(s):

Aleksandra Nestorovic ◽

Jovana Jasnic-Savovic ◽

Georgine Faulkner ◽

Dragica Radojkovic ◽

Snezana Kojic

Keyword(s):

Intracellular Signaling ◽

Pdz Domain ◽

Mechanical Stretch ◽

Regulatory Function ◽

Skeletal Muscle Differentiation ◽

Multiprotein Complex ◽

Zonula Occludens ◽

Binding Partners ◽

Zonula Occludens 1

The muscle ankyrin repeat protein Ankrd1 is localized in a mechanosensory complex of the sarcomeric I-band. It is involved in signaling pathways activated in response to mechanical stretch. It also acts as a transcriptional cofactor in the nucleus, playing an important role in cardiogenesis and skeletal muscle differentiation. To investigate its regulatory function in signaling we employed protein array methodology and identified 10 novel Ankrd1 binding partners among PDZ domain proteins known to act as platforms for multiprotein complex assembly. The zonula occludens protein-1 (ZO-1) was chosen for further analysis since its interaction with Ankrd2 had already been demonstrated. Both Ankrd2 and Ankrd1 have similar functions and localize in the same regions. We confirmed the interaction of Ankrd1 with ZO-1 protein and determined their subcellular distribution in HeLa cells, showing their colocalization in the cytoplasm. Our findings corroborate the role of Ankrd1 in intracellular signaling.

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The Second PDZ Domain of Zonula Occludens-1 Is Dispensable for Targeting to Connexin43 Gap Junctions

Cell Communication & Adhesion ◽

10.1080/15419060802014370 ◽

2008 ◽

Vol 15 (1-2) ◽

pp. 55-63 ◽

Cited By ~ 16

Author(s):

Andrew W. Hunter ◽

Robert G. Gourdie

Keyword(s):

Gap Junctions ◽

Pdz Domain ◽

Zonula Occludens ◽

Zonula Occludens 1 ◽

Connexin43 Gap Junctions

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Spatial Overlap of Claudin- and Phosphatidylinositol Phosphate-Binding Sites on the First PDZ Domain of Zonula Occludens 1 Studied by NMR

Molecules ◽

10.3390/molecules23102465 ◽

2018 ◽

Vol 23 (10) ◽

pp. 2465 ◽

Cited By ~ 3

Author(s):

Hidekazu Hiroaki ◽

Kaori Satomura ◽

Natsuko Goda ◽

Yukako Nakakura ◽

Minami Hiranuma ◽

...

Keyword(s):

Tight Junction ◽

Binding Site ◽

Solution Structure ◽

Pdz Domain ◽

Molecular Surface ◽

Head Group ◽

Functional Domain ◽

Phosphate Binding ◽

Zonula Occludens ◽

Zonula Occludens 1

Background: The tight junction is an intercellular adhesion complex composed of claudins (CLDs), occludin, and the scaffolding proteins zonula occludens 1 (ZO-1) and its two paralogs ZO-2 and ZO-3. ZO-1 is a multifunctional protein that contains three PSD95/Discs large/ZO-1(PDZ) domains. A key functional domain of ZO-1 is the first PDZ domain (ZO-1(PDZ1)) that recognizes the conserved C-termini of CLDs. Methods: In this study, we confirmed that phosphoinositides bound directly to ZO-1(PDZ1) by biochemical and solution NMR experiments. We further determined the solution structure of mouse ZO-1(PDZ1) by NMR and mapped the phosphoinositide binding site onto its molecular surface. Results: The phosphoinositide binding site was spatially overlapped with the CLD-binding site of ZO-1(PDZ1). Accordingly, inositol-hexaphosphate (phytic acid), an analog of the phosphoinositide head group, competed with ZO-1(PDZ)-CLD interaction. Conclusions: The results suggested that the PDZ domain–phosphoinositide interaction plays a regulatory role in biogenesis and homeostasis of the tight junction.

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Inhalational Anesthetics Disrupt Postsynaptic Density Protein-95, Drosophila Disc Large Tumor Suppressor, and Zonula Occludens-1 Domain Protein Interactions Critical to Action of Several Excitatory Receptor Channels Related to Anesthesia

Anesthesiology ◽

10.1097/aln.0000000000000609 ◽

2015 ◽

Vol 122 (4) ◽

pp. 776-786 ◽

Cited By ~ 10

Author(s):

Feng Tao ◽

Qiang Chen ◽

Yuko Sato ◽

John Skinner ◽

Pei Tang ◽

...

Keyword(s):

Protein Interactions ◽

Pdz Domain ◽

Postsynaptic Density ◽

Protein Protein Interactions ◽

Large Tumor ◽

Inhaled Anesthetics ◽

Zonula Occludens ◽

Peptide Binding Groove ◽

Binding Groove ◽

Postsynaptic Density Protein

Abstract Background: The authors have shown previously that inhaled anesthetics disrupt the interaction between the second postsynaptic density protein-95, Drosophila disc large tumor suppressor, and zonula occludens-1 (PDZ) domain of postsynaptic density protein-95 (PSD-95) and the C-terminus of N-methyl-d-aspartate receptor subunits NR2A and NR2B. The study data indicate that PDZ domains may serve as a molecular target for inhaled anesthetics. However, the underlying molecular mechanisms remain to be illustrated. Methods: Glutathione S-transferase pull-down assay, coimmunoprecipitation, and yeast two-hybrid analysis were used to assess PDZ domain–mediated protein–protein interactions in different conditions. Nuclear magnetic resonance spectroscopy was used to investigate isoflurane-induced chemical shift changes in the PDZ1–3 domains of PSD-95. A surface plasmon resonance–based BIAcore (Sweden) assay was used to examine the ability of isoflurane to inhibit the PDZ domain–mediated protein–protein interactions in real time. Results: Halothane and isoflurane dose-dependently inhibited PDZ domain–mediated interactions between PSD-95 and Shaker-type potassium channel Kv1.4 and between α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor subunit GluA2 and its interacting proteins—glutamate receptor–interacting protein or protein interacting with c kinase 1. However, halothane and isoflurane had no effect on PDZ domain–mediated interactions between γ-aminobutyric acid type B receptor and its interacting proteins. The inhaled anesthetic isoflurane mostly affected the residues close to or in the peptide-binding groove of PSD-95 PDZ1 and PDZ2 (especially PDZ2), while barely affecting the peptide-binding groove of PSD-95 PDZ3. Conclusion: These results suggest that inhaled anesthetics interfere with PDZ domain–mediated protein–protein interactions at several receptors important to neuronal excitation, anesthesia, and pain processing.

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