Artificial Polypeptide Scaffold for Protein Immobilization

An artificial polypeptide scaffold composed of surface anchor and protein capture domains was designed and expressed in vivo. By using a mutant E. coli phenylalanyl−tRNA synthetase, the photoreactive amino acid para-azidophenylalanine was incorporated into the surface anchor domain. Octyltrichlorosilane-treated surfaces were functionalized with this polypeptide by spin coating and photocrosslinking. The resulting protein films were shown to immobilize recombinant proteins through association of coiled coil heterodimer

Engineering Cooperativity in Biomotor-Protein Assemblies

A biosynthetic approach was developed to control and probe cooperativity in multiunit biomotor assemblies by linking molecular motors to artificial protein scaffolds. This approach provides precise control over spatial and elastic coupling between motors. Cooperative interactions between monomeric kinesin-1 motors attached to protein scaffolds enhance hydrolysis activity and microtubule gliding velocity. However, these interactions are not influenced by changes in the elastic properties of the scaffold, distinguishing multimotor transport from that powered by unorganized monomeric motors. These results highlight the role of supramolecular architecture in determining mechanisms of collective transport

Generation of Surface-Bound Multicomponent Protein Gradients

Spatial control of bioactive ligands is achieved by integrating microfluidics and protein engineering. The proteins of interest are mixed in a gradient generator and immobilized on artificial polypeptide scaffolds through the strong association of heterodimeric ZE/ZR leucine zipper pairs. Protein densities and gradient shapes are easily controlled and varied in this method

ErbB2 pY ‐1248 as a predictive biomarker for Parkinson's disease based on research with RPPA technology and in vivo verification

Aims: This study aims to reveal a promising biomarker for Parkinson's disease (PD) based on research with reverse phase protein array (RPPA) technology for the first time and in vivo verification, which gains time for early intervention in PD, thus increasing the effectiveness of treatment and reducing disease morbidity. Methods and Results: We employed RPPA technology which can assess both total and post‐translationally modified proteins to identify biomarker candidates of PD in a cellular PD model. As a result, the phosphorylation (pY‐1248) of the epidermal growth factor receptor (EGFR) ErbB2 is a promising biomarker candidate for PD. In addition, lapatinib, an ErbB2 tyrosine kinase inhibitor, was used to verify this PD biomarker candidate in vivo. We found that lapatinib‐attenuated dopaminergic neuron loss and PD‐like behavior in the zebrafish PD model. Accordingly, the expression of ErbB2pY‐1248 significantly increased in the MPTP‐induced mouse PD model. Our results suggest that ErbB2pY‐1248 is a predictive biomarker for PD. Conclusions: In this study, we found that ErbB2pY‐1248 is a predictive biomarker of PD by using RPPA technology and in vivo verification. It offers a new perspective on PD diagnosing and treatment, which will be essential in identifying individuals at risk of PD. In addition, this study provides new ideas for digging into biomarkers of other neurodegenerative diseases

Pomegranate Flower Water Extract Modulates AHR/BNIP3 to Improve Hepatic Insulin Signaling in Diabetic Mice

Objective: To investigate the effects and mechanisms of pomegranate flower water extract on hepatic insulin signaling in type 2 diabetic mice. Methods: C57BL/6J was randomly divided into normal group, model group, metformin group (Met), pomegranate flower water extract low-dose group (PFWL) and pomegranate flower water extract high-dose group (PFWH). The drug was administered continuously for 11 weeks. Mice were tested for body mass, fasting blood glucose (FBG), insulin (INS), triglycerides (TG), total cholesterol (TC) and insulin resistance index (HOMA-IR) was calculated. Hematoxylin-eosin (HE) staining was used to observe the pathologic changes in hepatic tissue. The expression levels of insulin receptor substrate 1 (IRS1), p-IRS1 (Ser307), protein kinase B (AKT), p-AKT (Ser473), glycogen synthase kinase-3β (Gsk3β), p-Gsk3β (S9), aromatidic hydrocarbon receptor (AhR), phosphatidylethanolamine N-methyltransferase (PEMT), and Bcl-2/adenovirus E1B19-kDa interacting protein 3 (BNIP3) in mouse liver tissues were determined by Western blot. Results: Compared with the model group, the FBG, INS, HOMA-IR, TG and TC levels were significantly decreased in the PFWH group (P<0.01). Intracellular fat droplets were significantly decreased in the liver of mice in the PFWH group. Western blot results showed that compared with the model group, IRS1, p-AKT (Ser473)/AKT, p-Gsk3β (S9)/Gsk3β, and BNIP3 protein expression were significantly increased in the liver of the PFWH group (P<0.01)，and p-IRS1 (Ser307)/IRS1, AHR, and PEMT protein expression were significantly decreased (P<0.01). Conclusion: PFW may inhibit hepatic lipid deposition by modulating AHR/BNIP3, and improve p-IRS1(Ser307)/p-AKT(Ser473)/p-GSK3β(S9) insulin signaling pathway transduction