Molecular mechanism of the susceptibility difference between HLA-B*27:02/04/05 and HLA-B*27:06/09 to ankylosing spondylitis: substitution analysis, MD simulation, QSAR modelling, and <i>in vitro</i> assay

<p>The human leukocyte antigen HLA-B27 is directly involved in the disease pathogenesis of ankylosing spondylitis (AS). HLA-B27 has a high degree of genetic polymorphism, with 105 currently known subtypes; the presence of aspartic acid at residue 116 (Asp116) has been found to play an essential role in AS susceptibility. Here, we systematically investigated the molecular mechanism of the susceptibility difference between the AS-associated subtypes HLA-B*27:02/04/05 and AS-unassociated subtypes HLA-B*27:06/09 to AS at sequence, structure, energetic and dynamic levels. In total seven variable residues were identified among the five studied HLA-B27 subtypes, in which Asp116 can be largely stabilized by a spatially vicinal, positively charged His114 through a salt bridge, while five other variable residues seem to have only a marginal effect on AS susceptibility. We also employed a quantitative structure–activity relationship approach to model the statistical correlation between peptide structure and affinity to HLA-B*27:05, a genetic ancestor of all other HLA-B27 subtypes and associated strongly with AS. The built regression predictor was verified rigorously through both internal cross-validation and external blind validation, and was then employed to identify potential HLA-B*27:05 binders from >20,000 cartilage-derived self-peptides. Subsequently, the binding potency of the top five antigenic peptides to HLA-B*27:05 was assayed <i>in vitro</i> using a FACS-based MHC stabilization experiment. Consequently, two (QRVGSDEFK and LRGAGTNEK) out of the five peptides were determined to have high affinity (BL₅₀ = 5.5 and 15.8 nM, respectively) and, as expected, both of them possess positively charged Lys at the C-terminus.</p

Highly Active and Stable CeO₂‑Promoted CuCl₂/Al₂O₃ Oxychlorination Catalysts Developed by Rational Design Using a Rate Diagram of the Catalytic Cycle

In this study, we have developed a method to predict the steady-state rate and Cu oxidation state during ethylene oxychlorination from a reaction rate diagram of the individual steps involved in the catalytic oxychlorination cycle. The steady state of the redox cycle is represented by a cross point of the reaction rates of the reduction and oxidation steps as a function of the Cu²⁺ in the rate diagram. Transient kinetics of elementary reactions and steady-state kinetics of the overall catalytic cycle were investigated in an operando study using combined mass and UV–vis-NIR spectrophotometry. The catalytic consequence of the promoters was then evaluated in terms of reduction and oxidation activity as well as number of active sites, site activity, and the catalyst oxidation state at steady state. Results revealed that the neat CuCl₂ catalysts operated at low Cu²⁺ at the steady-state conditions with stoichiometric feed composition, as a result of relatively low oxidation rate of Cu¹⁺. As a consequence of a high content of Cu¹⁺, ethylene conversion and selectivity are low, and the catalyst deactivates rapidly. By the promotion of the CuCl₂ catalyst by K, the reactor operates at a high Cu²⁺ concentration with much improved stability as a result of enhanced oxidation rate, but the catalyst has low activity due to significantly reduced reduction rate. Therefore, the rate diagram has been applied as a tool for a rational design of the CuCl₂-based oxychlorination catalysts, and Ce was proposed as the promoter due to its high promotion to the oxidation and low reactivity with Cu ions. It was found that the activity of the Ce-promoted catalysts increased 8 times compared to the neat CuCl₂ catalyst and moreover significantly improved the stability for the oxychlorination catalyst at steady state, due to the enhancement of both the rates of the reduction and oxidation. It is anticipated that the methodology developed here paves the way for a general method for catalyst design of heterogeneous catalysts where the catalyst undergoes oxidation state changes, in particular in redox reactions

Disruption of the Class IIa HDAC Corepressor Complex Increases Energy Expenditure and Lipid Oxidation

Drugs that recapitulate aspects of the exercise adaptive response have the potential to provide better treatment for diseases associated with physical inactivity. We previously observed reduced skeletal muscle class IIa HDAC (histone deacetylase) transcriptional repressive activity during exercise. Here, we find that exercise-like adaptations are induced by skeletal muscle expression of class IIa HDAC mutants that cannot form a corepressor complex. Adaptations include increased metabolic gene expression, mitochondrial capacity, and lipid oxidation. An existing HDAC inhibitor, Scriptaid, had similar phenotypic effects through disruption of the class IIa HDAC corepressor complex. Acute Scriptaid administration to mice increased the expression of metabolic genes, which required an intact class IIa HDAC corepressor complex. Chronic Scriptaid administration increased exercise capacity, whole-body energy expenditure and lipid oxidation, and reduced fasting blood lipids and glucose. Therefore, compounds that disrupt class IIa HDAC function could be used to enhance metabolic health in chronic diseases driven by physical inactivity