Lei Gao, Xiang Qiu, Jun Yang, Kangdelong Hu, Peilin Li, Wei Li, Feng Gao, Fabrice Gallou, Florian Kleinbeck, Xiaoguang Lei*
SCIENCE，29 JAN 2026 ，VOL 391, ISSUE 6784，DOI: 10.1126/SCIENCE.ADW336
Amide bond formation is widely used in pharmaceutical synthesis, typically involving stoichiometric coupling reagents to activate carboxylic acid substrates for a condensation reaction. As an alternative approach, we repurposed aldehyde dehydrogenases into oxidative amidases by creating a more hydrophobic and spacious catalytic pocket for amines to capture the thioester intermediate. This biocatalyst efficiently facilitates the formation of amide bonds between diverse aldehydes and amines. We also developed a two-step enzymatic cascade to synthesize amides from broadly available aliphatic alcohols. This biocatalytic strategy enabled the redesign of synthetic routes for five drug molecules. Our findings highlight the potential of oxidative amidases in advancing the synthesis of structurally diverse drug molecules through efficient amide bond formation
Structure and mechanism of the human bile acid transporter OSTα–OSTβ
Ke Wang, Junping Fan, Huiwen Chen, Bo Huang, Cheng Chi, Rui Yan, Di Wu, Feng Zhou, Wenhua Zhang, Juquan Jiang, Xiaoguang Lei& Daohua Jiang
https://doi.org/10.1038/s41586-025-09934-8
Bile acids (BAs) are crucial amphipathic surfactants that function as multifaceted regulators in various physiological processes, including nutrient absorption and distribution, lipid metabolism and inflammation. The human organic solute transporter αβ (OSTα–OSTβ; hereafter referred to as OSTα/β) is a BA transporter that has a key role in the secretion and distribution of BAs. Pathogenic mutations in OSTα/β have been associated with cholestasis. Despite the functional importance of OSTα/β in BA homeostasis, the stoichiometry and assembly of the complex and the molecular mechanism that underlies BA transport by OSTα/β remain unknown. Here we present cryo-electron microscopy structures of human OSTα/β in complex with cholesterols and an endogenous substrate, elucidating the structural basis for the function of OSTα/β. OSTα/β is assembled in a novel dimer-of-heterodimers manner: two OSTα units form the homodimeric core, with two OSTβ units bound to the periphery. OSTα adopts the G-protein-coupled-receptor (GPCR) fold and contains a unique cysteine-rich loop with seven palmitoylation sites; these cooperate with transmembrane helices 5 and 6, constituting a BA recognition site. A positive cavity in OSTα connects the BA site and facilitates the transmembrane translocation of BAs through OSTα/β. Together, this study reveals the architecture and transport mechanism of OSTα/β and provides insights into the structure–function relationships of this crucial transporter in BA homeostasis.
Cancer immunotherapies have revolutionized cancer treatment, yet many patients fail to respond. Activating innate immunity offers a promising approach to enhance therapeutic efficacy, but the signaling kinases directly regulating this process to boost antitumor responses remain elusive. Here we conduct an in vivo kinome CRISPR screen and identify CDK10 as a key suppressor of tumor immune surveillance. Mechanistically, CDK10 phosphorylates DNMT1 and RAP80 to reduce the accumulation of double-stranded RNA and R-loops, which alleviates the activation of innate immune pathways mediated by MDA5 and cGAS. Kinase inhibitor screens identify NVP-AST487 and ponatinib as selective CDK10 inhibitors. Both genetic and pharmacological inhibition of CDK10 activates MDA5 and cGAS pathways, fostering an immunoactive tumor microenvironment that enhances cancer immunotherapy in multiple mouse tumor models. Clinically, low CDK10 expression in tumors correlates with better immunotherapy responses. These findings establish CDK10 as a pivotal modulator of tumor immunity and a potential therapeutic target.
Mengze Sun , Baiyan Lu, Ying Yang, Junping Fan, Weiwei Ren, Xiaonan Chu, Yihui Gao, Jun Wu, Jue Wang, Han Ke, Zhiwen Liu, Shaojun Dai, Xiaoguang Lei*, and Chao Li*,
PNAS2025,122 (45),e2515322122
https://doi.org/10.1073/pnas.2515322122
Since its initial identification as the receptor for Rapid Alkalinization Factor 1 (RALF1), FERONIA (FER) receptor kinase has emerged as a central signaling hub coordi nating plant development, stress adaptation, and immune responses. Nevertheless, fundamental questions persist regarding the precise mechanisms of FER-mediated signal transduction and its context-dependent functional specialization in multicel lular processes. Here, we develop Ferovicin (FRV), a small-molecule inhibitor that specifically disrupts FER kinase activity, thereby enabling mechanistic dissection of FER. Cocrystallization and mutational analysis show that FRV selectively binds to the ATP-binding pocket of the kinase domain of FER and inhibits its kinase activity. Assisted by the FRV tool and quantitative phosphoproteomics, we characterized a series of signaling pathways and networks regulated by RALF1 and FER. Notably, our analysis reveals that RALF1 activates FER through phosphorylation at Ser695, which subsequently inhibits H+ -ATPase1/2 via phosphorylation at Ser899. This mechanism leads to apoplastic alkalinization and regulates cell expansion in the root meristem. Given the conservation of FRV binding sites in FER proteins across land plant species, FRV will serve as a valuable tool for dissecting FER signaling mechanisms as well as facilitating agricultural applications.
Arginine, a critical amino acid for protein structure and function, is involved in enzyme catalysis and macromolecular interactions. However, selectively targeting its reactive guanidine group has been challenging. Here, we utilized a probe, AP-1, based on phenylglyoxal, which demonstrated remarkable chemical selectivity and reactivity toward arginine residues. Using activity-based protein profifiling (ABPP), we explored the human proteome across four cancer cell lines, obtaining quantitative data for approximately 17 000 arginine residues. This analysis led to the identifification of several previously unreported hyperreactive arginine residues, including R43 of PKM, R171 of LDHA, R172 of LDHB, R341 of CKB, R168 of EIF4A1, and R118 of FUBP1, which are crucial for protein function. Notably, the mutation of CKB’s R341 inhibited cell proliferation and migration by downregulating energy supply. We also introduced ArGO-LDHA-1, a covalent inhibitor targeting LDHA’s hyperreactive arginine residues, showing potential to enhance chemotherapy effificacy. This work highlights the biological signifificance of arginine residues and provides a platform for large-scale profifiling of arginine reactivity.
Berberine bridge enzyme (BBE)-like enzymes catalyze various oxidative cyclization and dehydrogenation reactions in natural product biosynthesis, but the molecular mechanism underlying the selectivity remains unknown. Here, we elucidated the catalytic mechanism of BBE-like oxidases from Morus alba involved in the oxidative cyclization and dehydrogenation of moracin C. X-ray crystal structures of a functionally promiscuous flavin adenine dinucleotide (FAD)–bound oxidase, MaDS1, with and without an oxidative dehydrogenation product were determined at 2.03 Å and 2.21 Å resolution, respectively. Structure-guided mutagenesis and sequence analysis have identified a conserved aspartic acid that directs the reaction toward the oxidative dehydrogenation pathway. A combination of density functional theory (DFT) calculations and molecular dynamics (MD) simulations has revealed that aspartic acid acts as the catalytic base to deprotonate the carbon-cation intermediate to generate the dehydrogenated product, which otherwise undergoes a spontaneous 6π electrocyclization in the oxidative cyclization pathway to furnish the 2H-benzopyran product.
Dongshan Wu, Sanshan Wang, Haowen Zhang, Han Ke, Zeying Sun, Shuhan Xie, Yihui Gao, Jun Yang, Bingwu Wang, and Xiaoguang Lei*
J. Am. Chem. Soc. 2025, doi:10.1021/jacs.5c05656
Due to the invaluable properties of organofluorine compounds, incorporating a fluorinated unit has become necessary in pharmaceuticals, agrochemicals, and materials. However, achieving asymmetric fluorination such as trifluoromethylation through chemo- or biocatalysis has been a synthetic challenge. Here, we introduce a unique cooperative photoenzymatic catalysis for the enantioselective fluoroalkylation/cyclization cascade. This method, utilizing the engineered flavin-dependent “ene”-reductases (EREDs) and an exogenous photocatalyst (PC), produces a variety of fluorinated cyclic ketones with high yield and enantioselectivity. The discovery of stereocomplementary enzymes that provide access to both enantiomers of the cyclized products further enhances the synthetic applications of our method. The radical-trapping, spectroscopic, and kinetic studies have substantiated the interaction mode between the PC and the enzyme and demonstrated a cascade reaction mechanism involving a unique intermolecular addition of fluorinated radicals and a stereocontrolled intramolecular cyclization. Isotopic labeling experiments support flavin as the source of the hydrogen atom. Molecular dynamics simulations reveal that the binding interaction of the enzyme and the intermediate triggers the photoinduced enantioselective cyclization. This work underscores the potential of enzymes for the asymmetric synthesis of fluorinated compounds.
Dongshan Wu,† Zeying Sun,† Sanshan Wang, Jun Yang, Jingyuan He, and Xiaoguang Lei*
JACS Au 2025, doi:10.1021/jacsau.5c00633
Organic nitriles are significant in pharmaceuticals, agrochemicals, cosmetics, and materials. Although numerous cyanidation methods have been developed, more eco-friendly and green protocols for manufacturing alkyl nitriles are in high demand. Here, we report a photoenzymatic enantioselective intermolecular hydrocyanoalkylation of alkenes catalyzed by flavin-dependent “ene”-reductases. The discovery of stereocomplementary enzymes that provide access to both enantiomers of the high-value nitriles further showcases the synthetic applications of this method. Radical trapping, isotopic labeling, and spectroscopic experiments have elucidated the formation of a charge transfer complex at the protein active site. The single-electron reduction of the cyanoalkyl radical precursor by flavin hydroquinone yields a cyanoalkyl radical, which then undergoes intermolecular radical addition. This active site can stereoselectively control the radical-terminating hydrogen atom transfer, enabling the synthesis of enantioenriched γ-stereogenic nitriles. This work further expands the reactivity repertoire of biocatalytic transformations via non-natural radical mechanisms.
Auxin regulates many aspects of plant growth and development, featuring polar auxin transport mediated by auxin efflux and influx carriers. AUX1 is the prominent auxin importer that actively takes up natural and synthetic auxins. However, the precise mechanisms by which AUX1 recognizes and transports auxin remain elusive. Here, we describe the cryo-electron microscopy structures of Arabidopsis thaliana AUX1 in apo and auxin-bound forms, revealing the structural basis for auxin recognition. AUX1 assumes the LeuT-like fold in an inward conformation. The auxin analogue 2,4-D is recognized by polar residues in the middle of AUX1. We identify a putative cation site in AUX1, which plays a role in stabilizing the inward-facing conformation. His249 undergoes a large conformational shift, and mutation of it completely abolished transport activity, suggesting a crucial role of His249 in AUX1 gating. Together, this study provides a structural foundation for a deeper comprehension of auxin influx by AUX1-like carriers.
Xiaojiao Guo, Lei Gao, Shiwei Li, Jing Gao, Yuanyuan Wang, Jing Lv, Jiayi Wei, Jing Yang, Han Ke, Qi Ding, Jun Yang, Fusheng Guo, Haowen Zhang, Xiaoguang Lei* & Le Kang*
Nature (2025). DOI： 10.1038/s41586-025-09110-y
Aggregation pheromone, 4-vinylanisole (4VA), is specifically released by gregarious migratory locusts, and is crucial in forming locust swarms that cause destructive plagues1. Control of locust plagues relies heavily on the extensive application of chemical pesticides, which has led to severe environmental and health issues2. As pheromones are primary mediators of insect communication and behaviour3, exploring their biosynthesis can provide important cues to develop innovative behavioural regulators, potentially reducing the reliance on chemical pesticides. Here we resolve the biosynthesis of 4VA and behavioural responses of locusts when enzymes in the 4VA biosynthetic pathway are manipulated. The process initiates with phenylalanine derived from food plants and proceeds through three precursors: cinnamic acid, p-hydroxycinnamic acid and 4-vinylphenol (4VP). Notably, the conversion from 4VP to 4VA through methylation is unique to gregarious locusts. This step is catalysed by two crucial methyltransferases, 4VPMT1 and 4VPMT2. Guided by the X-ray co-crystal structure of 4VPMT2 bound with 4VP and S-adenosyl-L-methionine, we developed 4-nitrophenol as a substrate surrogate. We identified several chemicals that can block 4VA production by inhibiting the enzymatic activities of 4VPMT proteins, thereby suppressing locust aggregative behaviour. The findings uncover the chemical logic behind 4VA biosynthesis and pinpoint two crucial enzymes as novel targets for locust swarm management.
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Engineered aldehyde dehydrogenases for amide bond formation
Lei Gao, Xiang Qiu, Jun Yang, Kangdelong Hu, Peilin Li, Wei Li, Feng Gao, Fabrice Gallou, Florian Kleinbeck, Xiaoguang Lei*
SCIENCE，29 JAN 2026 ，VOL 391, ISSUE 6784，DOI: 10.1126/SCIENCE.ADW336
Amide bond formation is widely used in pharmaceutical synthesis, typically involving stoichiometric coupling reagents to activate carboxylic acid substrates for a condensation reaction. As an alternative approach, we repurposed aldehyde dehydrogenases into oxidative amidases by creating a more hydrophobic and spacious catalytic pocket for amines to capture the thioester intermediate. This biocatalyst efficiently facilitates the formation of amide bonds between diverse aldehydes and amines. We also developed a two-step enzymatic cascade to synthesize amides from broadly available aliphatic alcohols. This biocatalytic strategy enabled the redesign of synthetic routes for five drug molecules. Our findings highlight the potential of oxidative amidases in advancing the synthesis of structurally diverse drug molecules through efficient amide bond formation
Structure and mechanism of the human bile acid transporter OSTα–OSTβ
Structure and mechanism of the human bile acid transporter OSTα–OSTβ
Ke Wang, Junping Fan, Huiwen Chen, Bo Huang, Cheng Chi, Rui Yan, Di Wu, Feng Zhou, Wenhua Zhang, Juquan Jiang, Xiaoguang Lei& Daohua Jiang
https://doi.org/10.1038/s41586-025-09934-8
Bile acids (BAs) are crucial amphipathic surfactants that function as multifaceted regulators in various physiological processes, including nutrient absorption and distribution, lipid metabolism and inflammation. The human organic solute transporter αβ (OSTα–OSTβ; hereafter referred to as OSTα/β) is a BA transporter that has a key role in the secretion and distribution of BAs. Pathogenic mutations in OSTα/β have been associated with cholestasis. Despite the functional importance of OSTα/β in BA homeostasis, the stoichiometry and assembly of the complex and the molecular mechanism that underlies BA transport by OSTα/β remain unknown. Here we present cryo-electron microscopy structures of human OSTα/β in complex with cholesterols and an endogenous substrate, elucidating the structural basis for the function of OSTα/β. OSTα/β is assembled in a novel dimer-of-heterodimers manner: two OSTα units form the homodimeric core, with two OSTβ units bound to the periphery. OSTα adopts the G-protein-coupled-receptor (GPCR) fold and contains a unique cysteine-rich loop with seven palmitoylation sites; these cooperate with transmembrane helices 5 and 6, constituting a BA recognition site. A positive cavity in OSTα connects the BA site and facilitates the transmembrane translocation of BAs through OSTα/β. Together, this study reveals the architecture and transport mechanism of OSTα/β and provides insights into the structure–function relationships of this crucial transporter in BA homeostasis.
CDK10 suppresses nucleic acid sensors-mediated antitumor immunity
Gaoshan Xu, Fusheng Guo, Chuan He, Xiyong Wang, Bolin Xiang, Lifang Fan, Baoxiang Chen, Jiakun Peng, Yishuang Sun, Jie Shi, Xixin Xing, Yingmeng Yao, Panpan Dai, Haiou Li, Wenjun Xiong, Hudan Liu, Rui Xiao, Guoliang Qing, Congqing Jiang, Baishan Jiang, Xiaoguang Lei* & Jinfang Zhang*
Nature Cancer (2026)
https://doi.org/10.1038/s43018-025-01100-3
Cancer immunotherapies have revolutionized cancer treatment, yet many patients fail to respond. Activating innate immunity offers a promising approach to enhance therapeutic efficacy, but the signaling kinases directly regulating this process to boost antitumor responses remain elusive. Here we conduct an in vivo kinome CRISPR screen and identify CDK10 as a key suppressor of tumor immune surveillance. Mechanistically, CDK10 phosphorylates DNMT1 and RAP80 to reduce the accumulation of double-stranded RNA and R-loops, which alleviates the activation of innate immune pathways mediated by MDA5 and cGAS. Kinase inhibitor screens identify NVP-AST487 and ponatinib as selective CDK10 inhibitors. Both genetic and pharmacological inhibition of CDK10 activates MDA5 and cGAS pathways, fostering an immunoactive tumor microenvironment that enhances cancer immunotherapy in multiple mouse tumor models. Clinically, low CDK10 expression in tumors correlates with better immunotherapy responses. These findings establish CDK10 as a pivotal modulator of tumor immunity and a potential therapeutic target.
Unveiling FERONIA receptor kinase–mediated cellular mechanisms with a small-molecule inhibitor
Mengze Sun , Baiyan Lu, Ying Yang, Junping Fan, Weiwei Ren, Xiaonan Chu, Yihui Gao, Jun Wu, Jue Wang, Han Ke, Zhiwen Liu, Shaojun Dai, Xiaoguang Lei*, and Chao Li*,
PNAS2025,122 (45),e2515322122
https://doi.org/10.1073/pnas.2515322122
Since its initial identification as the receptor for Rapid Alkalinization Factor 1 (RALF1), FERONIA (FER) receptor kinase has emerged as a central signaling hub coordi nating plant development, stress adaptation, and immune responses. Nevertheless, fundamental questions persist regarding the precise mechanisms of FER-mediated signal transduction and its context-dependent functional specialization in multicel lular processes. Here, we develop Ferovicin (FRV), a small-molecule inhibitor that specifically disrupts FER kinase activity, thereby enabling mechanistic dissection of FER. Cocrystallization and mutational analysis show that FRV selectively binds to the ATP-binding pocket of the kinase domain of FER and inhibits its kinase activity. Assisted by the FRV tool and quantitative phosphoproteomics, we characterized a series of signaling pathways and networks regulated by RALF1 and FER. Notably, our analysis reveals that RALF1 activates FER through phosphorylation at Ser695, which subsequently inhibits H+ -ATPase1/2 via phosphorylation at Ser899. This mechanism leads to apoplastic alkalinization and regulates cell expansion in the root meristem. Given the conservation of FRV binding sites in FER proteins across land plant species, FRV will serve as a valuable tool for dissecting FER signaling mechanisms as well as facilitating agricultural applications.
Quantitative Reactivity Profiling of Functional Arginine Residues in Human Cancer Cell Line Proteomes
Wenbo Zhao+, Yuliang Tang+, Yihui Gao, Qi Ding, Qiang Li, Wenyang Li, and Xiaoguang Lei*
Angew. Chem. Int. Ed. 2025, e202515603
Arginine, a critical amino acid for protein structure and function, is involved in enzyme catalysis and macromolecular interactions. However, selectively targeting its reactive guanidine group has been challenging. Here, we utilized a probe, AP-1, based on phenylglyoxal, which demonstrated remarkable chemical selectivity and reactivity toward arginine residues. Using activity-based protein profifiling (ABPP), we explored the human proteome across four cancer cell lines, obtaining quantitative data for approximately 17 000 arginine residues. This analysis led to the identifification of several previously unreported hyperreactive arginine residues, including R43 of PKM, R171 of LDHA, R172 of LDHB, R341 of CKB, R168 of EIF4A1, and R118 of FUBP1, which are crucial for protein function. Notably, the mutation of CKB’s R341 inhibited cell proliferation and migration by downregulating energy supply. We also introduced ArGO-LDHA-1, a covalent inhibitor targeting LDHA’s hyperreactive arginine residues, showing potential to
enhance chemotherapy effificacy. This work highlights the biological signifificance of arginine residues and provides a platform for large-scale profifiling of arginine reactivity.
Aspartic acid residues in BBE-like enzymes from Morus alba promote a function shift from oxidative cyclization to dehydrogenation
Nianxin Guo, Jun Gu, Qingyang Zhou, Fang Liu, Haoran Dong, Qi Ding, Qixuan Wang, Dongshan Wu, Jun Yang, Junping Fan, Lei Gao*, Kendall N. Houk*, and Xiaoguang Lei*
PNAS, 2025, 122 (34) e2504346122. DOI: 10.1073/pnas.2504346122
Berberine bridge enzyme (BBE)-like enzymes catalyze various oxidative cyclization and dehydrogenation reactions in natural product biosynthesis, but the molecular mechanism underlying the selectivity remains unknown. Here, we elucidated the catalytic mechanism of BBE-like oxidases from Morus alba involved in the oxidative cyclization and dehydrogenation of moracin C. X-ray crystal structures of a functionally promiscuous flavin adenine dinucleotide (FAD)–bound oxidase, MaDS1, with and without an oxidative dehydrogenation product were determined at 2.03 Å and 2.21 Å resolution, respectively. Structure-guided mutagenesis and sequence analysis have identified a conserved aspartic acid that directs the reaction toward the oxidative dehydrogenation pathway. A combination of density functional theory (DFT) calculations and molecular dynamics (MD) simulations has revealed that aspartic acid acts as the catalytic base to deprotonate the carbon-cation intermediate to generate the dehydrogenated product, which otherwise undergoes a spontaneous 6π electrocyclization in the oxidative cyclization pathway to furnish the 2H-benzopyran product.
Cooperative Photoenzymatic Catalysis for Enantioselective Fluoroalkylation/Cyclization Cascade
Dongshan Wu, Sanshan Wang, Haowen Zhang, Han Ke, Zeying Sun, Shuhan Xie, Yihui Gao, Jun Yang,
Bingwu Wang, and Xiaoguang Lei*
J. Am. Chem. Soc. 2025, doi:10.1021/jacs.5c05656
Due to the invaluable properties of organofluorine compounds, incorporating a fluorinated unit has become necessary in pharmaceuticals, agrochemicals, and materials. However, achieving asymmetric fluorination such as trifluoromethylation through chemo- or biocatalysis has been a synthetic challenge. Here, we introduce a unique cooperative photoenzymatic catalysis for the enantioselective fluoroalkylation/cyclization cascade. This method, utilizing the engineered flavin-dependent “ene”-reductases (EREDs) and an exogenous photocatalyst (PC), produces a variety of fluorinated cyclic ketones with high yield and enantioselectivity. The discovery of stereocomplementary enzymes that provide access to both enantiomers of the cyclized products further enhances the synthetic applications of our method. The radical-trapping, spectroscopic, and kinetic studies have substantiated the interaction mode between the PC and the enzyme and demonstrated a cascade reaction mechanism involving a unique intermolecular addition of fluorinated radicals and a stereocontrolled intramolecular cyclization. Isotopic labeling experiments support flavin as the source of the hydrogen atom. Molecular dynamics simulations reveal that the binding interaction of the enzyme and the intermediate triggers the photoinduced enantioselective cyclization. This work underscores the potential of enzymes for the asymmetric synthesis of fluorinated compounds.
Enantioselective Radical Hydrocyanoalkylation of Alkenes via Photoenzymatic Catalysis
Dongshan Wu,† Zeying Sun,† Sanshan Wang, Jun Yang, Jingyuan He, and Xiaoguang Lei*
JACS Au 2025, doi:10.1021/jacsau.5c00633
Organic nitriles are significant in pharmaceuticals, agrochemicals, cosmetics, and materials. Although numerous cyanidation methods have been developed, more eco-friendly and green protocols for manufacturing alkyl nitriles are in high demand. Here, we report a photoenzymatic enantioselective intermolecular hydrocyanoalkylation of alkenes catalyzed by flavin-dependent “ene”-reductases. The discovery of stereocomplementary enzymes that provide access to both enantiomers of the high-value nitriles further showcases the synthetic applications of this method. Radical trapping, isotopic labeling, and spectroscopic experiments have elucidated the formation of a charge transfer complex at the protein active site. The single-electron reduction of the cyanoalkyl radical precursor by flavin hydroquinone yields a cyanoalkyl radical, which then undergoes intermolecular radical addition. This active site can stereoselectively control the radical-terminating hydrogen atom transfer, enabling the synthesis of enantioenriched γ-stereogenic nitriles. This work further expands the reactivity repertoire of biocatalytic transformations via non-natural radical mechanisms.
Structural basis of auxin recognition and transport in plant influx carrier AUX1
Huiwen Chen1, Junping Fan1, Cheng Chi1, Jun Zhao, Di Wu, Xiaoguang Lei*, Xingwang Deng*, Daohua Jiang*
Molecular Plant, 2025.
doi.org/10.1016/j.molp.2025.06.015
Auxin regulates many aspects of plant growth and development, featuring polar auxin transport mediated by auxin efflux and influx carriers. AUX1 is the prominent auxin importer that actively takes up natural and synthetic auxins. However, the precise mechanisms by which AUX1 recognizes and transports auxin remain elusive. Here, we describe the cryo-electron microscopy structures of Arabidopsis thaliana AUX1 in apo and auxin-bound forms, revealing the structural basis for auxin recognition. AUX1 assumes the LeuT-like fold in an inward conformation. The auxin analogue 2,4-D is recognized by polar residues in the middle of AUX1. We identify a putative cation site in AUX1, which plays a role in stabilizing the inward-facing conformation. His249 undergoes a large conformational shift, and mutation of it completely abolished transport activity, suggesting a crucial role of His249 in AUX1 gating. Together, this study provides a structural foundation for a deeper comprehension of auxin influx by AUX1-like carriers.
Decoding 4-vinylanisole biosynthesis and pivotal enzymes in locusts
Xiaojiao Guo, Lei Gao, Shiwei Li, Jing Gao, Yuanyuan Wang, Jing Lv, Jiayi Wei, Jing Yang, Han Ke, Qi Ding, Jun Yang, Fusheng Guo, Haowen Zhang, Xiaoguang Lei* & Le Kang*
Nature (2025). DOI： 10.1038/s41586-025-09110-y
Aggregation pheromone, 4-vinylanisole (4VA), is specifically released by gregarious migratory locusts, and is crucial in forming locust swarms that cause destructive plagues1. Control of locust plagues relies heavily on the extensive application of chemical pesticides, which has led to severe environmental and health issues2. As pheromones are primary mediators of insect communication and behaviour3, exploring their biosynthesis can provide important cues to develop innovative behavioural regulators, potentially reducing the reliance on chemical pesticides. Here we resolve the biosynthesis of 4VA and behavioural responses of locusts when enzymes in the 4VA biosynthetic pathway are manipulated. The process initiates with phenylalanine derived from food plants and proceeds through three precursors: cinnamic acid, p-hydroxycinnamic acid and 4-vinylphenol (4VP). Notably, the conversion from 4VP to 4VA through methylation is unique to gregarious locusts. This step is catalysed by two crucial methyltransferases, 4VPMT1 and 4VPMT2. Guided by the X-ray co-crystal structure of 4VPMT2 bound with 4VP and S-adenosyl-L-methionine, we developed 4-nitrophenol as a substrate surrogate. We identified several chemicals that can block 4VA production by inhibiting the enzymatic activities of 4VPMT proteins, thereby suppressing locust aggregative behaviour. The findings uncover the chemical logic behind 4VA biosynthesis and pinpoint two crucial enzymes as novel targets for locust swarm management.