Structural Basis for the Interaction of 14-3-3<i>β</i>with Tricarboxylic Acid Cycle Intermediate Malate

The protein family of 14-3-3(s) has risen to a position of higher importance as an adaptor protein in cell biology. The seven highly conserved human 14-3-3 proteins coordinate diverse cellular processes including apoptosis, DNA damage response, protein trafficking, and others. In liver hepatocytes, 14-3-3β binds to Ser196-phosphorilated glucose-responsive carbohydrate response element-binding protein (ChREBP) to inhibit converting excess carbohydrate to fat by regulating the nuclear/cytosol trafficking of ChREBP. Here, we report X-ray crystal structures of homodimeric mammalian 14-3-3β in its apo, Malate-bound forms. The determined apo structure was captured with one monomer in the closed state, whereas the other one had an open conformation. Strikingly, 14-3-3β binds Malate dynamically with a double-closed state, which is distinct from all previously characterized 14-3-3(s) and target ligand-binding modes. Malate docks into a first-time observed cofactor pocket located at the concaved interface of 14-3-3β helices α2, α3, α4 through mainly electrostatic and hydrogen interactions. Such a Tricarboxylic Acid Cycle intermediate Malate bond model might offer a new approach to further analyze insulin-independent 14-3-3/ChREBP pathway of de novo fat synthesis in the liver.

Optimization of high-pressure processing-assisted extraction of pectic polysaccharides from three berries

Background and objectives:High-pressure processing(HPP)is a promising assistive method to extract pectic polysaccharides with high rhamnogalacturonan I(RG-I)domain and berries are sources of such pectic polysaccharides.This study extracts pectic polysaccharides from goji berry,raspberry,and cranberry,examines how HPP influences the pectic polysaccharide structure of three berries,and provides a basis for the extraction and modification of pectic polysaccharides with specific structure and bioactivity.Materials and methods:An orthogonal test was performed to optimize the HPP-assisted alkali method to extract the high yield and high RG-I content pectic polysaccharides from three berries.Structural information of pectic polysaccharides extracted by the HPP method and conventional methods were compared from the perspectives of monosaccharide composition,molecular weight,Fourier transform infrared spectroscopy,andnuclearmagneticresonancespectroscopy.Results:For raspberry,the optimal conditions consisted of a pressure of 50o MPa,a pH of 13,and a pressure-holding time of 12 min,while the optimal conditions for goji berry and cranberry were both 40o MPa,pH 13,and 15 min.Under the optimal conditions,the yields for goji berry,raspberry,and cranberry were 10.49%,16.63%,and 17.52%,respectively,and RG-l contents were 81.85%,83.30%,and 63.22%,respectively.HPP showed an effect to degrade homogalacturonan backbones and side chains and increase the RG-I content to some extent.Conclusion:HPP-assisted alkali method was revealed to be an efficient method to extract high RG-I content pectic polysaccharides,especially for cranberry,and was a potential method to modify pectic polysaccharide structure in a certain way.