Recruitment of ubiquitin E2 enzymes is determined jointly by the U-box domains and substrates of E3 ligases.

Ubiquitination is a cascade reaction involving E1, E2, and E3 enzymes. The orthogonal ubiquitin transfer (OUT) method has been previously established to identify potential substrates of E3 ligases. In this study, we verified the ubiquitination of five substrates mediated by the E3 ligases CHIP and E4B. To further explore the activity of U-box domains of E3 ligases, two mutants with the U-box domains interchanged between CHIP and E4B were generated. They exhibited a significantly reduced ubiquitination ability. Additionally, different E3s recruited similar E2 ubiquitin-conjugating enzymes when ubiquitinating the same substrates, highlighting that U-box domains determined the E2 recruitment, while the substrate determined the E2 selectivity. This study reveals the influence of substrates and U-box domains on E2 recruitment, providing a novel perspective on the function of U-box domains of E3 ligases.

CHIP suppresses the proliferation and migration of A549 cells by mediating the ubiquitination of eIF2α and upregulation of tumor suppressor RBM5.

The protein kinase RNA-like endoplasmic reticulum kinase (PERK)-eukaryotic translation initiation factor 2 subunit α (eIF2α) pathway plays an essential role in endoplasmic reticulum (ER) stress. When the PERK-eIF2α pathway is activated, PERK phosphorylates eIF2α (p-eIF2α) at Ser51 and quenches global protein synthesis. In this study, we verified eIF2α as a bona fide substrate of the E3 ubiquitin ligase carboxyl terminus of the HSC70-interaction protein (CHIP) both in vitro and in cells. CHIP mediated the ubiquitination and degradation of nonphosphorylated eIF2α in a chaperone-independent manner and promoted the upregulation of the cyclic AMP-dependent transcription factor under endoplasmic reticulum stress conditions. Cyclic AMP-dependent transcription factor induced the transcriptional enhancement of the tumor suppressor genes PTEN and RBM5. Although transcription was enhanced, the PTEN protein was subsequently degraded by CHIP, but the expression of the RBM5 protein was upregulated, thereby suppressing the proliferation and migration of A549 cells. Overall, our study established a new mechanism that deepened the understanding of the PERK-eIF2α pathway through the ubiquitination and degradation of eIF2α. The crosstalk between the phosphorylation and ubiquitination of eIF2α shed light on a new perspective for tumor progression.

Longitudinal and transverse photophoretic force on a homogeneous sphere exerted by a Bessel beam with selective polarizations.

Predicting the photophoretic force exerted on an optical absorptive particle in a gaseous medium is a challenging problem because the problems of electromagnetic scattering, heat transfer, and gaseous molecule dynamics are involved and coupled with each other. Based on the calculation of the source function distribution inside a homogeneous sphere excited by a Bessel beam using the generalized Lorenz-Mie theory, analytical expressions of the asymmetry vector, which is the key quantity in the calculation of photophoretic force, are given using the adjoint boundary value method. Numerical simulations are performed to analyze the influences of polarization, the half-cone angle, and the beam order of the incident beam, particle size, and absorptivity of the particle on the asymmetry vector for both on-axis and off-axis illuminations. Longitudinal and transverse photophoretic forces on a homogeneous sphere are displayed for the slip-flow regime of gaseous media. The results offer important insights into the working mechanism underpinning the development of heat-mediated optical manipulation techniques and the measurement of the refractive index of particles.

Microcompression Method for Determining the Size-Dependent Elastic Properties of PMMA Microcapsules Containing n-Octadecane.

Accurate evaluation of the shell elastic modulus of microcapsules is of great significance to understanding their performance during production, processing, and applications. In this work, microcompression was employed to investigate the elastic behaviors of a single microcapsule. It was modeled as a microsphere with a core-shell structure compressed between two rigid plates. Based on the assumption that the contact pressure between the microsphere and plates obeys parabolic distribution, a microcompression method derived from the Reissner's theory and the modified Hertz contact theory was established to evaluate the shell elastic modulus. Applications were carried out on poly(methylmethacrylate) (PMMA) microcapsules containing n-octadecane. The average elastic modulus of PMMA shells measured by the proposed microcompression method agrees well with that of the bulk PMMA sample. Furthermore, the elastic modulus of PMMA shells was found to have size dependence on the diameter of the microcapsules. Finally, finite element models combined with the newly proposed method were constructed to accurately predict the microcompression behaviors of microcapsules with different sizes.

Potential solution processible phosphorescent iridium complexes toward applications in doped light-emitting diodes: rapid syntheses and optical and morphological characterizations.

A series of carbazol-9-yl end-capped red-emitting heteroleptic iridium complexes was readily synthesized using post Suzuki coupling. The appealing solubility, photoluminescent characteristics, and morphological stability enabled the current heteroleptic iridium complexes to be highly desirable phosphorescent dopant emitters in doped light-emitting diodes for the purpose of resisting molecular aggregations in the doped emitting layers.