Advanced glycation end products (AGEs), a heterogeneous compound existed in processed foods, are related to chronic diseases when they are accumulated excessively in human organs. Protein-bound Nε-(carboxymethyl) lysine (CML) as a typical AGE, is widely determined to evaluate AGEs level in foods and in vivo. This study investigated the intestinal absorption of three protein-bound CML originated from main food raw materials (soybean, wheat and peanut). After in vitro gastrointestinal digestion, the three protein-bound CML digests were ultrafiltered and divided into four fractions: less than 1 kDa, between 1 and 3 kDa, between 3 and 5 kDa, greater than 5 kDa. Caco-2 cell monolayer model was further used to evaluate the intestinal absorption of these components. Results showed that the absorption rates of soybean protein isolate (SPI)-, glutenin (Glu)-, peanut protein isolate (PPI)-bound CML were 30.18%, 31.57% and 29.5%, respectively. The absorption rates of components with MW less than 5 kDa accounted for 19.91% (SPI-bound CML), 22.59% (Glu-bound CML), 23.64% (PPI-bound CML), respectively, and these samples were absorbed by paracellular route, transcytosis route and active route via PepT-1. Taken together, these findings demonstrated that all three protein-bound CML digests with different MW can be absorbed in diverse absorption pathways by Caco-2 cell monolayer model. This research provided a theoretical basis for scientific evaluation of digestion and absorption of AGEs in food.

Protein post-translational modifications (PTMs) are crucial for cancer cells to adapt to hypoxia; however, the functional significance of lysine crotonylation (Kcr) in hypoxia remains unclear. Herein we report a quantitative proteomics analysis of global crotonylome under normoxia and hypoxia, and demonstrate 128 Kcr site alterations across 101 proteins in MDA-MB231 cells. Specifically, we observe a significant decrease in K131cr, K156cr and K220cr of phosphoglycerate kinase 1 (PGK1) upon hypoxia. Enoyl-CoA hydratase 1 (ECHS1) is upregulated and interacts with PGK1, leading to the downregulation of PGK1 Kcr under hypoxia. Abolishment of PGK1 Kcr promotes glycolysis and suppresses mitochondrial pyruvate metabolism by activating pyruvate dehydrogenase kinase 1 (PDHK1). A low PGK1 K131cr level is correlated with malignancy and poor prognosis of breast cancer. Our findings show that PGK1 Kcr is a signal in coordinating glycolysis and the tricarboxylic acid (TCA) cycle and may serve as a diagnostic indicator for breast cancer.

Quorum sensing (QS) can regulate the production of multiple functional factors in bacteria, but the process of identifying its regulatory targets is very complex and labor-intensive. In this study, an efficient and rapid method to find QS targets through prediction was used. The genome of Lactiplantibacillus plantarum (L. plantarum) L3 was sequenced and characterized, and then linked the L. plantarum L3 genome to the STRING database for QS system regulatory target prediction. A total of 3,167,484 base pairs (bps) were examined from the genome of L. plantarum L3, and 30 QS-related genes were discovered (including luxS). The STRING database prediction indicated that the 30 QS-related genes are mainly involved in the regulation of nine metabolic pathways. Furthermore, metE, metK, aroB, cysE, and birA1 were predicted to be regulatory targets of the LuxS/AI-2 QS system, and these five targets were validated based on quantitative real-time PCR and content determination. Successful elucidation of the LuxS/AI-2 QS system\'s key targets and regulation mechanism in L. plantarum L3 demonstrated the effectiveness of the new approach for predicting QS targets and provides a scientific basis for future work on improving regulation of functional factor production.

Lactate-derived histone lactylation is involved in multiple pathological processes through transcriptional regulation. The role of lactate-derived histone lactylation in the repair of spinal cord injury (SCI) remains unclear. Here we report that overall lactate levels and lactylation are upregulated in the spinal cord after SCI. Notably, H4K12la was significantly elevated in the microglia of the injured spinal cord, whereas exogenous lactate treatment further elevated H4K12la in microglia after SCI. Functionally, lactate treatment promoted microglial proliferation, scar formation, axon regeneration, and locomotor function recovery after SCI. Mechanically, lactate-mediated H4K12la elevation promoted PD-1 transcription in microglia, thereby facilitating SCI repair. Furthermore, a series of rescue experiments confirmed that a PD-1 inhibitor or microglia-specific AAV-sh-PD-1 significantly reversed the therapeutic effects of lactate following SCI. This study illustrates the function and mechanism of lactate/H4K12la/PD-1 signaling in microglia-mediated tissue repair and provides a novel target for SCI therapy.

Histone lysine crotonylation, an evolutionarily conserved modification differing from acetylation, exerts pivotal control over diverse biological processes. Among these are gene transcriptional regulation, spermatogenesis, and cell cycle processes. However, the dynamic changes and functions of histone crotonylation in preimplantation embryonic development in mammals remain unclear. Here, we show that the transcription coactivator P300 functions as a writer of histone crotonylation during embryonic development. Depletion of P300 results in significant developmental defects and dysregulation of the transcriptome of embryos. Importantly, we demonstrate that P300 catalyzes the crotonylation of histone, directly stimulating transcription and regulating gene expression, thereby ensuring successful progression of embryo development up to the blastocyst stage. Moreover, the modification of histone H3 lysine 18 crotonylation (H3K18cr) is primarily localized to active promoter regions. This modification serves as a distinctive epigenetic indicator of crucial transcriptional regulators, facilitating the activation of gene transcription. Together, our results propose a model wherein P300-mediated histone crotonylation plays a crucial role in regulating the fate of embryonic development.

The identification of readers, an important class of proteins that recognize modified residues at specific sites, is essential to uncover the biological roles of post-translational modifications. Photoreactive crosslinkers are powerful tools for investigating readers. However, existing methods usually employ synthetically challenging photoreactive warheads, and their high-energy intermediates generated upon irradiation, such as nitrene and carbene, may cause substantial non-specific crosslinking. Here we report dimethylsulfonium as a methyllysine mimic that binds to specific readers and subsequently crosslinks to a conserved tryptophan inside the binding pocket through single-electron transfer under ultraviolet irradiation. The crosslinking relies on a protein-templated σ-π electron donor-acceptor interaction between sulfonium and indole, ensuring excellent site selectivity for tryptophan in the active site and orthogonality to other methyllysine readers. This method could escalate the discovery of methyllysine readers from complex cell samples. Furthermore, this photo crosslinking strategy could be extended to develop other types of microenvironment-dependent conjugations to site-specific tryptophan.

This study aimed to investigate the effect of lysine (Lys) on quality profile promotion and saltiness compensation in reduced‑sodium salt frankfurters. The results showed that the cooking loss of reduced‑sodium salt frankfurters decreased from 5.63 to 3.45% when the Lys level increased from 0.1 to 0.7%, as well as enhanced water contents and saltiness in a Lys dose-dependent manner. Moreover, the corresponding peak area percentage (A21) of reduced‑sodium salt frankfurters remarkably increased from 79.63% to 81.48% with the increased level of Lys. However, a higher level of Lys (≥ 0.5%) obviously reduced the textural properties of reduced‑sodium salt frankfurters, which was clearly verified by looser and rougher microstructures. Furthermore, hydrogen bonds were found to be the dominant molecular force in Lys-added reduced‑sodium salt frankfurters. Meanwhile, 0.3% Lys was found to exhibit the optimal sodium salt-replacing effect due to the highest degree of quality profile promotion and saltiness compensation. Additionally, directional triangle sensory evaluation further confirmed that the reduced‑sodium salt frankfurters treated with 0.3% Lys was perceived to be similar to control frankfurters, resulting a 50% salt-reduction effect. Thus, our results suggested that Lys could be applied as an efficient sodium salt alternative in reduced‑sodium salt frankfurters.

Lysine plays a crucial role in promoting development, enhancing immune function, and improving the function of central nervous system tissues. The two configurational isomers of amino acids have significantly different effects. Currently, methods for chiral recognition of lysine have been reported; however, previous detection methods have drawbacks such as expensive equipment and complicated detection processes. Fluorescence analysis, on the other hand, boasts high sensitivity, strong selectivity, and simple operation. In this study, we synthesized four novel Binaphthyl-Amine (BINAM)-based fluorescent probes capable of specifically identifying the L-configuration of lysine among the twenty amino acids that constitute human proteins. The enantiomeric fluorescence enhancement ratio (ef or ΔIL/ΔID) reached up to 15.29, demonstrating high enantioselectivity. In addition, we assessed the probe\'s recognition capabilities under varying pH levels, reaction times, and metal ion conditions, along with its limit of detection (LOD) and quantum yield. Our results suggest that this probe serves as a highly stable tool for the detection of chiral lysine.

Glucose and lipid metabolism are essential energy sources for the body. Dysregulation in these metabolic pathways is a significant risk factor for numerous acute and chronic diseases, including type 2 diabetes (T2DM), Alzheimer\'s disease (AD), obesity, and cancer. Post-translational modifications (PTMs), which regulate protein structure, localization, function, and activity, play a crucial role in managing cellular glucose and lipid metabolism. Among these PTMs, lysine methylation stands out as a key dynamic modification vital for the epigenetic regulation of gene transcription. Emerging evidence indicates that lysine methylation significantly impacts glucose and lipid metabolism by modifying key enzymes and proteins. This review summarizes the current understanding of lysine methylation\'s role and regulatory mechanisms in glucose and lipid metabolism. We highlight the involvement of methyltransferases (KMTs) and demethylases (KDMs) in generating abnormal methylation signals affecting these metabolic pathways. Additionally, we discuss the chemical biology and pharmacology of KMT and KDM inhibitors and targeted protein degraders, emphasizing their clinical implications for diseases such as diabetes, obesity, neurodegenerative disorders, and cancers. This review suggests that targeting lysine methylation in glucose and lipid metabolism could be an ideal therapeutic strategy for treating these diseases.

This study aimed to investigate the effects of dietary crude protein (CP) and lysine levels on growth performance, slaughter performance, meat quality, and myofiber characteristics of slow-growing chicken. A 3 × 3 factorial experiment was arranged, and the chickens were fed with 3 levels of dietary CP (16.0%, 17.0%, 18.0%) and 3 levels of dietary lysine (0.69%, 0.84%, 0.99%). A total of 540 8-week-old Beijing-You Chicken (BYC) female growing chickens were randomly allocated to 9 groups, 5 replicates per group, and 12 chickens per replicate. The birds were randomly allocated to one of the 9 experimental diets. Growth performance, slaughter performance, meat quality, and myofiber characteristics were determined at 16 weeks of age. The results showed that dietary CP level and the interaction of dietary CP and lysine levels affected average feed intake (AFI) (p < 0.05). The AFI in the 16.0% CP and 17.0% CP groups was higher than in the 18.0% CP group (p < 0.05). Dietary CP levels significantly affected body weight gain (BWG) (p < 0.05) at 9 to 16 weeks. The 18.0% CP group had the highest BWG (93.99 g). Dietary CP levels affected the percentage of leg muscle yield, and the percentage of leg muscle yield of the 16.0% CP group was significantly lower than that in the other two groups (p < 0.05). Dietary CP and lysine levels alone and their interactions did not affect pH24h, drip loss, and cooking loss of breast muscle (p > 0.05). The shear force of the 18.0% CP group (29.55 N) was higher than that in the other two groups (p < 0.01). Dietary CP level affected myofiber characteristic (p < 0.01), with the lowest myofiber density (846.35 p·mm-2) and the largest myofiber diameter (30.92 μm) at 18.0% CP level. Dietary lysine level affected myofiber diameter, endomysium thickness, perimysium thickness (p < 0.01), with the largest myofiber diameter (29.29 μm) obtained at 0.84% lysine level, the largest endomysium thickness (4.58 μm) at 0.69% lysine level, and the largest perimysium thickness (9.26 μm) at 0.99% lysine level. Myofiber density was negatively correlated with myofiber diameter and endomysium thickness (R = -0.883, R = -0.523, p < 0.01); perimysium thickness had a significant negative correlation with shear force (R = -0.682, p < 0.05). Therefore, reducing dietary CP level and adding appropriate lysine can reduce myofiber diameter and increase perimysium thickness, reducing shear force and improving meat tenderness. A high lysine level (0.99%) in the low-CP (16.0%) diet can improve meat tenderness by regulating the myofiber characteristic without affecting production performance.