Emerging Role of Nicotinamide Riboside in Health and Diseases

Among all the NAD+ precursors, nicotinamide riboside (NR) has gained the most attention as a potent NAD+-enhancement agent. This recently discovered vitamin, B3, has demonstrated excellent safety and efficacy profiles and is orally bioavailable in humans. Boosting intracellular NAD+ concentrations using NR has been shown to provide protective effects against a broad spectrum of pathological conditions, such as neurodegenerative diseases, diabetes, and hearing loss. In this review, an integrated overview of NR research will be presented. The role NR plays in the NAD+ biosynthetic pathway will be introduced, followed by a discussion on the synthesis of NR using chemical and enzymatic approaches. NR’s effects on regulating normal physiology and pathophysiology will also be presented, focusing on the studies published in the last five years

Substrate-Dependent Sensitivity of SIRT1 to Nicotinamide Inhibition

SIRT1 is the most extensively studied human sirtuin with a broad spectrum of endogenous targets. It has been implicated in the regulation of a myriad of cellular events, such as gene transcription, mitochondria biogenesis, insulin secretion as well as glucose and lipid metabolism. From a mechanistic perspective, nicotinamide (NAM), a byproduct of a sirtuin-catalyzed reaction, reverses a reaction intermediate to regenerate NAD+ through “base exchange”, leading to the inhibition of the forward deacetylation. NAM has been suggested as a universal sirtuin negative regulator. Sirtuins have evolved different strategies in response to NAM regulation. Here, we report the detailed kinetic analysis of SIRT1-catalyzed reactions using endogenous substrate-based synthetic peptides. A novel substrate-dependent sensitivity of SIRT1 to NAM inhibition was observed. Additionally, SIRT1 demonstrated pH-dependent deacetylation with normal solvent isotope effects (SIEs), consistent with proton transfer in the rate-limiting step. Base exchange, in contrast, was insensitive to pH changes with no apparent SIEs, indicative of lack of proton transfer in the rate-limiting step. Consequently, NAM inhibition was attenuated at a high pH in proteated buffers. Our study provides new evidence for “activation by de-repression” as an effective sirtuin activation strategy

Plasmodium falciparum Nicotinamidase as A Novel Antimalarial Target

Inhibition of Plasmodium falciparum nicotinamidase could represent a potential antimalarial since parasites require nicotinic acid to successfully recycle nicotinamide to NAD+, and importantly, humans lack this biosynthetic enzyme. Recently, mechanism-based inhibitors of nicotinamidase have been discovered. The most potent compound inhibits both recombinant P. falciparum nicotinamidase and parasites replication in infected human red blood cells (RBCs). These studies provide evidence for the importance of nicotinamide salvage through nicotinamidase as a central master player of NAD+ homeostasis in P. falciparum

Post-Translational Modifications and Diabetes

Diabetes and its associated complications have increasingly become major challenges for global healthcare. The current therapeutic strategies involve insulin replacement therapy for type 1 diabetes (T1D) and small-molecule drugs for type 2 diabetes (T2D). Despite these advances, the complex nature of diabetes necessitates innovative clinical interventions for effective treatment and complication prevention. Accumulative evidence suggests that protein post-translational modifications (PTMs), including glycosylation, phosphorylation, acetylation, and SUMOylation, play important roles in diabetes and its pathological consequences. Therefore, the investigation of these PTMs not only sheds important light on the mechanistic regulation of diabetes but also opens new avenues for targeted therapies. Here, we offer a comprehensive overview of the role of several PTMs in diabetes, focusing on the most recent advances in understanding their functions and regulatory mechanisms. Additionally, we summarize the pharmacological interventions targeting PTMs that have advanced into clinical trials for the treatment of diabetes. Current challenges and future perspectives are also provided

Changes of Water Migration and Texture Characteristics of Oil-tea Camellia Seeds during Fresh Storage

In order to clarify water migration and textural changes in oil-tea camellia seeds during fresh storage after harvest, low-field nuclear magnetic resonance and magnetic resonance imaging techniques were used to clarify moisture state and distribution law and textural profile analysis was used to monitor the changes in the texture characteristics taking the oil-tea camellia seeds produced in Guizhou Province as test materials. And correlation analysis was carried out. The results showed that the water content of the oil-tea camellia seeds decreased continuously during storage. The water content of Qianyu 1 decreased from 33.86%±3.03% (0 d) to 8.64%±0.24% (56 d), and that of Xianglin 210 decreased from 53.03%±3.36% (0 d) to 10.73%±0.25% (56 d). The rate of decrease in kernel water content was higher than that of the oil-tea camellia seeds. The proportion of non-flowable water was the highest in the oil-tea camellia seeds, whereas that in Qianyu 1 decreased to 62.89% and that in Xianglin 210 decreased to 60.64% after 56 d of storage. The hydrogen proton density image of the fresh oil-tea camellia seeds was bright. Local water loss increased as storage time was prolonged, and the image gradually approached the background color. The fracture and hardness properties of the oil-tea camellia seeds decreased continuously during storage, and that of the seed kernels changed flexural; The springiness of the seed kernels was gradually lost; The cohesiveness of Qianyu 1 changed slightly, while that of Xianglin 210 increased first and then decreased. Correlation analysis revealed a very extremely significant positive correlation between the water content and the total peak area of oil-tea camellia seeds (P<0.01), a significant positive correlation between fracture of oil-tea camellia seeds (P<0.05), and a very significant positive correlation between fracture and hardness (P<0.01). Additionally, a very significant positive correlation was observed between the kernel water content and kernel fracture, kernel hardness and kernel springiness (P<0.01). This study provides basic data for fresh oil-tea camellia seeds pressing technology

Development of Activity-Based Chemical Probes for Human Sirtuins

Sirtuins consume stoichiometric amounts of nicotinamide adenine dinucleotide (NAD⁺) to remove an acetyl group from lysine residues. These enzymes have been implicated in regulating various cellular events and have also been suggested to mediate the beneficial effects of calorie restriction (CR). However, controversies on sirtuin biology also peaked during the past few years because of conflicting results from different research groups. This is partly because these enzymes have been discovered recently and the intricate interaction loops between sirtuins and other proteins make the characterization of them extremely difficult. Current molecular biology and proteomics techniques report protein abundance rather than active sirtuin content. Innovative chemical tools that can directly probe the functional state of sirtuins are desperately needed. We have obtained a set of powerful activity-based chemical probes that are capable of assessing the active content of sirtuins in model systems. These probes consist of a chemical “warhead” that binds to the active site of active enzyme and a handle that can be used for the visualization of these enzymes by fluorescence. In complex native proteome, the probes can selectively “highlight” the active sirtuin components. Furthermore, these probes were also able to probe the dynamic change of sirtuin activity in response to cellular stimuli. These chemical probes and the labeling strategies will provide transformative technology to allow the direct linking of sirtuin activity to distinct physiological processes. They will create new opportunities to investigate how sirtuins provide health benefits in adapting cells to environmental cues and provide critical information to dissect sirtuin regulatory networks

Scalable Synthesis and Purification of Acetylated Phosphatidyl Choline Headgroup

The acetylated headgroup of the most abundant mammalian phospholipid, 1,2-diacetyl-3-<i>sn</i>-phosphatidyl choline (DAcPC), has several important applications in research. For instance, it can be dissolved in the same amount of water as in the fluid PC bilayer, to create a surrogate of a PC headgroup stratum, for studying the solvation of small molecules and the influence of their structure on the process. In contrast to PC derivatives with longer acyl chains, DAcPC does not self-aggregate, rendering the aqueous solution homogeneous and suitable for simplified analyses of interactions of molecules with the headgroups. Several studies have been published where DAcPC was used in a crudely purified form. Here we describe a one-step preparation of DAcPC from commercially available bulk chemicals and purification of the product by crystallization and washing. The process gives a good yield and is easily scalable. The availability of enantiopure, crystalline DAcPC could open the door to more extensive biochemical, pharmacological, and nutritional studies of this interesting chemical