Intracellular targeting, interaction with Src homology 3 (SH3) domains and rolipram-detected conformational switches in cAMP-specific PDE4A phosphodiesterase

1997 ◽  
Vol 25 (2) ◽  
pp. 374-381 ◽  
Author(s):  
M. D. Houslay ◽  
G. Scotland ◽  
S. Erdogan ◽  
E. Huston ◽  
S. Mackenzie ◽  
...  
Keyword(s):  
Sh3 Domains ◽  
Src Homology 3 ◽  
Intracellular Targeting ◽  
Src Homology ◽  
Conformational Switches

scholarly journals Interactions between cytosolic components of the NADPH oxidase: p40phox interacts with both p67phox and p47phox

1996 ◽  
Vol 317 (3) ◽  
pp. 919-924 ◽  
Author(s):  
Frans B. WIENTJES ◽  
George PANAYOTOU ◽  
Emer REEVES ◽  
Anthony W. SEGAL
Keyword(s):  
Nadph Oxidase ◽  
Binding Assays ◽  
Phagocytic Cells ◽  
Sh3 Domains ◽  
Cytosolic Proteins ◽  
Src Homology 3 ◽  
Vitro Binding ◽  
Src Homology ◽  
Flavocytochrome B

The NADPH oxidase of neutrophils and other bone-marrow-derived phagocytic cells is a multi-component system consisting of a flavocytochrome b in the plasma membrane and at least four cytosolic proteins. Three of the cytosolic proteins contain src homology 3 (SH3) domains, two each in p47phox and p67phox, and one in p40phox. All three translocate from the cytosol to the flavocytochrome in the membrane upon stimulation of the cells. A small G-protein, p21rac, is also involved in activation of the oxidase. The three cytosolic phox proteins occur as a complex in the cytosol and the strongest interaction appeared to be between p67phox and p40phox. We have investigated the interaction between p40phox and the other two cytosolic phox proteins by in vitro binding assays. An affinity-bead approach was used as well as a biosensor technique (surface plasmon resonance). We observed the strongest attachment between p40phox and p67phox where the binding was between the N-terminal half of p67phox and the C-terminal half of p40phox, and did not appear to involve SH3 domains and proline-rich sequences. p40phox also bound p47phox but more weakly than it did p67phox.

scholarly journals Capacity of simian immunodeficiency virus strain mac Nef for high-affinity Src homology 3 (SH3) binding revealed by ligand-tailored SH3 domains

2002 ◽  
Vol 83 (12) ◽  
pp. 3147-3152 ◽  
Author(s):  
Marita Hiipakka ◽  
Kalle Saksela
Keyword(s):  
Simian Immunodeficiency Virus ◽  
Binding Motif ◽  
Sh3 Domains ◽  
Src Homology 3 ◽  
High Affinity ◽  
Loop Region ◽  
Immunodeficiency Virus ◽  
Src Homology ◽  
Hiv 1

The simian immunodeficiency virus (SIV) Nef protein contains a consensus Src-homology 3 (SH3) binding motif. However, no SH3-domain proteins showing strong binding to SIV Nef have yet been found, and its potential capacity for high-affinity SH3 binding has therefore remained unproven. Here we have used phage-display-assisted protein engineering to develop artificial SH3 domains that bind tightly to SIV strain mac (SIVmac) Nef. Substitution of six amino acids in the RT loop region of Hck-SH3 with the sequence E/DGWWG resulted in SH3 domains that bound in vitro to SIVmac Nef much better than the natural Hck- or Fyn-SH3 domains. These novel SH3 domains also efficiently associated with SIVmac Nef when co-expressed in 293T cells and displayed a strikingly differential specificity when compared with SH3 domains similarly targeted for binding to human immunodeficiency virus type 1 (HIV-1) Nef. Thus, SIVmac Nef is competent for high-affinity SH3 binding, but its natural SH3 protein partners are likely to be different from those of HIV-1 Nef.

scholarly journals Large-Scale Screening of Preferred Interactions of Human Src Homology-3 (SH3) Domains Using Native Target Proteins as Affinity Ligands

2016 ◽  
Vol 15 (10) ◽  
pp. 3270-3281 ◽  
Author(s):  
Arunas Kazlauskas ◽  
Constanze Schmotz ◽  
Tapio Kesti ◽  
Jussi Hepojoki ◽  
Iivari Kleino ◽  
...  
Keyword(s):  
Large Scale ◽  
Sh3 Domains ◽  
Affinity Ligands ◽  
Target Proteins ◽  
Src Homology 3 ◽  
Src Homology ◽  
Large Scale Screening

Insights into the structure and protein–protein interactions of the pro-apoptotic protein ASPP2

2007 ◽  
Vol 35 (5) ◽  
pp. 966-969 ◽  
Author(s):  
S. Rotem ◽  
C. Katz ◽  
A. Friedler
Keyword(s):  
Intermolecular Interactions ◽  
Protein Interactions ◽  
Intramolecular Interaction ◽  
Protein Protein Interactions ◽  
Sh3 Domains ◽  
Src Homology 3 ◽  
Rich Domain ◽  
C Terminus ◽  
Apoptotic Protein ◽  
Src Homology

ASPP (apoptosis-stimulating protein of p53) 2 is a pro-apoptotic protein that stimulates the p53-mediated apoptotic response. Here, we provide an overview of the structure and protein–protein interactions of ASPP2. The C-terminus of ASPP2 contains Ank (ankyrin) repeats and an SH3 domain (Src homology 3 domain). The Ank–SH3 domains mediate interactions between ASPP2 and numerous proteins involved in apoptosis such as p53 and Bcl-2. The proline-rich domain of ASPP2 is unfolded in its native state, but was not shown to mediate intermolecular interactions. Instead, it makes an intramolecular domain–domain interaction with the Ank–SH3 C-terminal domains of ASPP2. This intramolecular interaction between the unstructured proline-rich domain and the structured Ank–SH3 domains in ASPP2, which is possible due to the unfolded nature of the proline-rich domain, is proposed to have an important role in regulating the intermolecular interactions of ASPP2 with its partner proteins.

scholarly journals Differential Recognition Preferences of the Three Src Homology 3 (SH3) Domains from the Adaptor CD2-associated Protein (CD2AP) and Direct Association with Ras and Rab Interactor 3 (RIN3)

2015 ◽  
Vol 290 (42) ◽  
pp. 25275-25292 ◽  
Author(s):  
Evgenia Rouka ◽  
Philip C. Simister ◽  
Melanie Janning ◽  
Joerg Kumbrink ◽  
Tassos Konstantinou ◽  
...  
Keyword(s):  
Sh3 Domains ◽  
Src Homology 3 ◽  
Differential Recognition ◽  
Direct Association ◽  
Src Homology

scholarly journals A high affinity RIM-binding protein/Aplip1 interaction prevents the formation of ectopic axonal active zones

eLife ◽  
2015 ◽  
Vol 4 ◽  
Author(s):  
Matthias Siebert ◽  
Mathias A Böhme ◽  
Jan H Driller ◽  
Husam Babikir ◽  
Malou M Mampell ◽  
...  
Keyword(s):  
Synaptic Vesicles ◽  
Protein Transport ◽  
Binding Protein ◽  
Sh3 Domains ◽  
Src Homology 3 ◽  
High Affinity ◽  
Pxxp Motif ◽  
Transport Mutants ◽  
Src Homology ◽  
Active Zones

Synaptic vesicles (SVs) fuse at active zones (AZs) covered by a protein scaffold, at Drosophila synapses comprised of ELKS family member Bruchpilot (BRP) and RIM-binding protein (RBP). We here demonstrate axonal co-transport of BRP and RBP using intravital live imaging, with both proteins co-accumulating in axonal aggregates of several transport mutants. RBP, via its C-terminal Src-homology 3 (SH3) domains, binds Aplip1/JIP1, a transport adaptor involved in kinesin-dependent SV transport. We show in atomic detail that RBP C-terminal SH3 domains bind a proline-rich (PxxP) motif of Aplip1/JIP1 with submicromolar affinity. Pointmutating this PxxP motif provoked formation of ectopic AZ-like structures at axonal membranes. Direct interactions between AZ proteins and transport adaptors seem to provide complex avidity and shield synaptic interaction surfaces of pre-assembled scaffold protein transport complexes, thus, favouring physiological synaptic AZ assembly over premature assembly at axonal membranes.

scholarly journals Activation of C-terminal Src kinase (Csk) by phosphorylation at serine-364 depends on the Csk-Src homology 3 domain

2003 ◽  
Vol 372 (1) ◽  
pp. 271-278 ◽  
Author(s):  
Sheraz YAQUB ◽  
Hilde ABRAHAMSEN ◽  
Bastian ZIMMERMAN ◽  
Natalya KHOLOD ◽  
Knut Martin TORGERSEN ◽  
...  
Keyword(s):  
Src Kinase ◽  
Kinase Domain ◽  
Structural Domain ◽  
Wild Type ◽  
Sh3 Domains ◽  
Src Homology 3 ◽  
Src Homology 3 Domain ◽  
Src Homology ◽  
Kinase A ◽  
Significant Activation

In the present study, we investigate the mechanism for the protein kinase A (PKA)-mediated activation of C-terminal Src kinase (Csk). Although isolated Csk kinase domain was phosphorylated at Ser364 by PKA to the same stoichiometry as wild-type Csk, significant activation of the isolated Csk kinase domain by PKA was observed only in the presence of the purified Src homology 3 domain (SH3 domain). Furthermore, the interaction between the SH3 and kinase domains was facilitated by PKA-mediated phosphorylation of the kinase domain, as evaluated by surface plasmon resonance. This suggests that an overall structural domain organization and interaction between the kinase and SH3 domains are important for the activity of Csk and its regulation by PKA.

scholarly journals Vinexin β Interacts with Hepatitis C Virus NS5A, Modulating Its Hyperphosphorylation To Regulate Viral Propagation

2015 ◽  
Vol 89 (14) ◽  
pp. 7385-7400 ◽  
Author(s):  
Wei Xiong ◽  
Jie Yang ◽  
Mingzhen Wang ◽  
Hailong Wang ◽  
Zhipeng Rao ◽  
...  
Keyword(s):  
Life Cycle ◽  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Nonstructural Protein ◽  
Casein Kinase ◽  
Genome Replication ◽  
Sh3 Domains ◽  
Content Type ◽  
Src Homology 3 ◽  
Src Homology

ABSTRACTHepatitis C virus (HCV) nonstructural protein 5A (NS5A) is essential for HCV genome replication and virion production and is involved in the regulation of multiple host signaling pathways. As a proline-rich protein, NS5A is capable of interacting with various host proteins containing Src homology 3 (SH3) domains. Previous studies have suggested that vinexin, a member of the sorbin homology (SoHo) adaptor family, might be a potential binding partner of NS5A by yeast two-hybrid screening. However, firm evidence for this interaction is lacking, and the significance of vinexin in the HCV life cycle remains unclear. In this study, we demonstrated that endogenously and exogenously expressed vinexin β coimmunoprecipitated with NS5A derived from different HCV genotypes. Two residues, tryptophan (W307) and tyrosine (Y325), in the third SH3 domain of vinexin β and conserved Pro-X-X-Pro-X-Arg motifs at the C terminus of NS5A were indispensable for the vinexin-NS5A interaction. Furthermore, downregulation of endogenous vinexin β significantly suppressed NS5A hyperphosphorylation and decreased HCV replication, which could be rescued by expressing a vinexin β short hairpin RNA-resistant mutant. We also found that vinexin β modulated the hyperphosphorylation of NS5A in a casein kinase 1α-dependent on manner. Taken together, our findings suggest that vinexin β modulates NS5A phosphorylation via its interaction with NS5A, thereby regulating HCV replication, implicating vinexin β in the viral life cycle.IMPORTANCEHepatitis C virus (HCV) nonstructural protein NS5A is a phosphoprotein, and its phosphorylation states are usually modulated by host kinases and other viral nonstructural elements. Additionally, cellular factors containing Src homology 3 (SH3) domains have been reported to interact with proline-rich regions of NS5A. However, it is unclear whether there are any relationships between NS5A phosphorylation and the NS5A-SH3 interaction, and little is known about the significance of this interaction in the HCV life cycle. In this work, we demonstrate that vinexin β modulates NS5A hyperphosphorylation through the NS5A-vinexin β interaction. Hyperphosphorylated NS5A induced by vinexin β is casein kinase 1α dependent and is also crucial for HCV propagation. Overall, our findings not only elucidate the relationships between NS5A phosphorylation and the NS5A-SH3 interaction but also shed new mechanistic insight onFlaviviridaeNS5A (NS5) phosphorylation. We believe that our results may afford the potential to offer an antiviral therapeutic strategy.

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