TRAF6-mediated ubiquitination of AKT in the nucleus is a critical event underlying the desensitization of G protein-coupled receptors.

BACKGROUND: Desensitization of G protein-coupled receptors (GPCRs) refers to the attenuation of receptor responsiveness by prolonged or intermittent exposure to agonists. The binding of ß-arrestin to the cytoplasmic cavity of the phosphorylated receptor, which competes with the G protein, has been widely accepted as an extensive model for explaining GPCRs desensitization. However, studies on various GPCRs, including dopamine D2-like receptors (D2R, D3R, D4R), have suggested the existence of other desensitization mechanisms. The present study employed D2R/D3R variants with different desensitization properties and utilized loss-of-function approaches to uncover the mechanisms underlying GPCRs homologous desensitization, focusing on the signaling cascade that regulates the ubiquitination of AKT. RESULTS: AKT undergoes K8/14 ubiquitination by TRAF6, which occurs in the nucleus and promotes its membrane recruitment, phosphorylation and activation under receptor desensitization conditions. The nuclear entry of TRAF6 relies on the presence of the importin complex. Src regulates the nuclear entry of TRAF6 by mediating the interaction between TRAF6 and importin ß1. Ubiquitinated AKT translocates to the plasma membrane where it associates with Mdm2 to phosphorylate it at the S166 and S186 residues. Thereafter, phosphorylated Mdm2 is recruited to the nucleus, resulting in the deubiquitination of ß-Arr2. The deubiquitinated ß-Arr2 then forms a complex with Gßγ, which serves as a biomarker for GPCRs desensitization. Like in D3R, ubiquitination of AKT is also involved in the desensitization of ß2 adrenoceptors. CONCLUSION: Our study proposed that the property of a receptor that causes a change in the subcellular localization of TRAF6 from the cytoplasm to the nucleus to mediate AKT ubiquitination could initiate the desensitization of GPCRs.

Generation of discrete higher-order optical vortex lattice at focus.

Higher-order vortices (HOVs) extend the dimensions of optical vortex regulation, which is of great significance in optical communication and optical tweezers. Herein, we demonstrate an alternative scheme to produce a HOV in the focus plane using multiple Laguerre-Gaussian (LG) beam interference, termed a discrete higher-order optical vortex lattice (DHOVL). The modulation depth of the DHOVL exceeds 2π. In this case, the topological charge (TC) of the DHOVL is determined by the difference of the phase period between the innermost and the outermost interference beams. Compared with a conventional HOV (CHOV), the vortex exists in a form of multiple unit singularities sharing a dark core. In addition, the average orbital angular momentum per photon of the DHOVL increases with increasing TC, surpassing that of the CHOV. This work provides a novel, to the best of our knowledge, scheme to produce a HOV, which will facilitate several advanced applications, including optical micromanipulation, optical sensing and imaging, and optical fabrication.

Switchable rotation of metal nanostructures in an intensity chirality-invariant focus field.

Light-induced rotation is a fundamental motion form that is of great significance for flexible and multifunctional manipulation modes. However, current optical rotation by a single optical field is mostly unidirectional, where switchable rotation manipulation is still challenging. To address this issue, we demonstrate a switchable rotation of non-spherical nanostructures within a single optical focus field. Interestingly, the intensity of the focus field is chiral invariant. The rotation switch is a result of the energy flux reversal in front and behind the focal plane. We quantitatively analyze the optical force exerted on a metal nanorod at different planes, as well as the surrounding energy flux. Our experimental results indicate that the direct switchover of rotational motion is achievable by adjusting the relative position of the nanostructure to the focal plane. This result enriches the basic motion mode of micro-manipulation and is expected to create potential opportunities in many application fields, such as biological cytology and optical micromachining.

All-optically controlled holographic plasmonic vortex array for multiple metallic particles manipulation.

Due to the sub-diffraction-limited size and giant field enhancement, plasmonic tweezers have a natural advantage in trapping metallic particles. However, the strict excitation condition makes it difficult to generate an arbitrary plasmonic field in a controllable manner, thus narrowing its practical applications. Here, we propose an all-optical plasmonic field shaping method based on a digital holographic algorithm and generate plasmonic vortex arrays with controllable spot numbers, spatial location, and topological charge. Our experimental results demonstrate that multiple gold particles can be stably trapped and synchronously rotated in the vortex arrays, and the particles' kinestate can be dynamically switched. The proposed holographic plasmonic vortex tweezers are suitable for a broadband particle trapping, and this method can be generalized to other surface electromagnetic waves like Bloch surface wave.

Ubiquitination of GRK2 Is Required for the ß-Arrestin-Biased Signaling Pathway of Dopamine D2 Receptors to Activate ERK Kinases.

A class-A GPCR dopamine D2 receptor (D2R) plays a critical role in the proper functioning of neuronal circuits through the downstream activation of both G-protein- and ß-arrestin-dependent signaling pathways. Understanding the signaling pathways downstream of D2R is critical for developing effective therapies with which to treat dopamine (DA)-related disorders such as Parkinson's disease and schizophrenia. Extensive studies have focused on the regulation of D2R-mediated extracellular-signal-regulated kinase (ERK) 1/2 signaling; however, the manner in which ERKs are activated upon the stimulation of a specific signaling pathway of D2R remains unclear. The present study conducted a variety of experimental techniques, including loss-of-function experiments, site-directed mutagenesis, and the determination of protein interactions, in order to investigate the mechanisms underlying ß-arrestin-biased signaling-pathway-mediated ERK activation. We found that the stimulation of the D2R ß-arrestin signaling pathway caused Mdm2, an E3 ubiquitin ligase, to move from the nucleus to the cytoplasm and interact with tyrosine phosphorylated G-protein-coupled receptor kinase 2 (GRK2), which was facilitated by Src, a non-receptor tyrosine kinase. This interaction led to the ubiquitination of GRK2, which then moved to the plasma membrane and interacted with activated D2R, followed by the phosphorylation of D2R as well as the mediation of ERK activation. In conclusion, Mdm2-mediated GRK2 ubiquitination, which is selectively triggered by the stimulation of the D2R ß-arrestin signaling pathway, is necessary for GRK2 membrane translocation and its interaction with D2R, which in turn mediates downstream ERK signaling. This study is primarily novel and provides essential information with which to better understand the detailed mechanisms of D2R-dependent signaling.

Time-varying orbital angular momentum in tight focusing of ultrafast pulses.

The orbital angular momentum (OAM) of light has important applications in a variety of fields, including optical communication, quantum information, super-resolution microscopic imaging, particle trapping, and others. However, the temporal properties of OAM in ultrafast pulses and in the evolution process of spin-orbit coupling has yet to be revealed. In this work, we theoretically studied the spatiotemporal property of time-varying OAM in the tightly focused field of ultrafast light pulses. The focusing of an incident light pulse composed of two time-delayed femtosecond sub-pulses with the same OAM but orthogonal spin states is investigated, and the ultrafast dynamicsa time delay of OAM variation during the focusing process driven by the spin-orbit coupling is visualized. Temporal properties of three typical examples, including formation, increase, and transformation of topological charge are investigated to reveal the non-uniform evolutions of phase singularities, local topological charges, self-torques, and time-varying OAM per photon. This work could deepen the understanding of spin-orbit coupling in time domain and promote many promising applications such as ultrafast OAM modulation, laser micromachining, high harmonic generation, and manipulation of molecules and nanostructures.

Energy flow inversion in an intensity-invariant focusing field.

Dependence of light intensity on energy flow is the most intuitive presentation of an optical field. This dependence, however, also limits the applications to the interaction of the light field with matter. For further insight into this, we demonstrate a novel case of the optical field, named as the counterintuitive chiral intensity field (CCIF), in the highly focusing situation: the energy flow reverses during the propagation but the intensity distribution pattern is kept approximately invariant. Our results show that, in this process, the mode correlation decreases rapidly while the intensity correlation remains invariant in the focus area. Furthermore, this property is still valid even if the pattern helicity and number of spiral arms are changed. This work deepens the understanding of the relationship between energy flow and field intensity, and it will offer diversified operations in many applications, such as optical micromanipulation, optical fabrication, etc.

Controllable propagation and transformation of chiral intensity field at focus.

As an intrinsic feature of the optical field, chirality could induce many novel phenomena due to the interaction between chiral light and matter. Thus, the generation of optical fields possessing 2D or 3D chiral intensity patterns, called chiral intensity fields, has been widely studied. However, the control of chiral intensity field along the optical axis is still a challenge. Here, we propose a method to manipulate the axial propagation property of a focused chiral intensity field. Two modulation effects are realized: extended chiral intensity field with a focal depth >2λ at 90% mode correlation and tunable transformation of chirality during the axial propagation. This method is simple, stable, and easy to perform and therefore offers broad applications especially in optical tweezers and metamaterial fabrication.

Grafted optical vortex with controllable orbital angular momentum distribution.

In an optical vortex (OV) field, the orbital angular momentum (OAM) distribution strongly depends on the intensity, which results in difficulty in OAM independent modulation. To overcome this limitation, we propose a grafted optical vortex (GOV) via spiral phase reconstruction of two or more OVs with different topological charges (TCs). To remain the annular shape of the GOV's intensity, the Dirac δ-function is employed to restrict the energy in a ring. Theoretical analysis and manipulation experiments of polystyrene microspheres show that the magnitude and direction of the GOV's local OAM are controllable by modulating the grafted TCs while the intensity remains constant. The results of this work provide an ingenious method to control the local tangential force on the light ring, which will promote potential applications in optical trapping and rotating micro-particles.

Optical vortex shaping via a phase jump factor.

Topological charge (TC) of an optical vortex (OV) is a crucial parameter. We propose two factors, namely, the phase jump factor and the phase gradient factor, to replace the parameter of TC through unwrapping the TC definition integral. Based on these two factors, we report on a novel OV, referred to as the remainder-phase optical vortex (ROV). The properties of the ROV are studied in depth by adjusting these two factors. Results show that the phase gradient factor determines the total orbital angular momentum (OAM), whereas the phase jump factor decides the number of split unit vortices and reshapes the structure of the OAM distribution. This work provides a novel OV with controllable OAM distribution, which will open up new applications such as particle manipulation, beam shaping, and micro-fabrication.