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1.
J Periodontol ; 2023 Nov 01.
Artículo en Inglés | MEDLINE | ID: mdl-37910482

RESUMEN

BACKGROUND: Antimicrobial photodynamic therapy (aPDT) is an effective method for eradicating bacteria in periodontal therapy. Standard aPDT requires the insertion of a laser tip into a periodontal pocket, in which the direction of irradiation is limited. Therefore, we devised an aPDT method that uses a transgingival near-infrared wavelength and indocyanine green-encapsulated and chitosan-coated nanoparticles as a photosensitizer. METHODS: Forty patients undergoing supportive periodontal therapy, who had a single root tooth with a pocket of 5 mm or deeper, were used as subjects. In the test group, aPDT was performed by laser irradiation from outside the gingiva using photosensitizer nanoparticles. In the control group, pseudo aPDT without photosensitizer was performed by transgingival irradiation. Subgingival plaque was sampled from inside the pocket before, immediately after, and 1 week after treatment, and evaluated by colony counting and real-time polymerase chain reaction. RESULTS: There were no significant differences in age, sex, periodontal pocket depth, and bleeding on probing between the test and control groups. Compared with the colony count before treatment, the count in the test group was significantly reduced immediately after treatment. The number of patients with colony reduction to ≤50% and ≤10% was significantly higher in the test group than in the control group. None of the participants reported pain, although one participant reported discomfort. CONCLUSION: As a bacterial control method for residual pockets in patients undergoing supportive periodontal therapy, transgingival aPDT is a promising treatment strategy that is not generally accompanied by pain or discomfort.

2.
BMC Microbiol ; 23(1): 285, 2023 10 05.
Artículo en Inglés | MEDLINE | ID: mdl-37798648

RESUMEN

BACKGROUND: Previous studies have revealed a nitric oxide (NO) metabolic cycle in which NO, nitrate (NO3-), and nitrite (NO2-) circulate. The NO produced in this cycle serves as a signalling molecule that regulates actinorhodin (ACT) production via the DevS/DevR NO-dependent two-component system (TCS) in Streptomyces coelicolor A3(2) M145. However, the mechanisms involved in the regulation of NO signalling in S. coelicolor have not yet been elucidated. Mycothiol (MSH), a thiol molecule produced by Actinomyces, is involved in the defence mechanisms against oxidative stress. Therefore, this study focused on the correlation between intracellular NO and MSH levels. RESULTS: To investigate the interaction of MSH with endogenously produced NO, we generated an S. coelicolor A3(2) strain deficient in MSH biosynthesis. This mutant strain exhibited a decrease in low-molecular-weight S-nitrosothiols and intracellular NO levels during culture compared to those of the wild-type strain. Moreover, the mutant strain exhibited reduced activity of the DevS/DevR TCS, a regulator of NO homeostasis and ACT production, from the early stage of culture, along with a decrease in ACT production compared to those of the wild-type strain. CONCLUSIONS: This study suggests that MSH maintains intracellular NO homeostasis by forming S-nitrosomycothiol, which induces NO signalling. Finally, we propose a metabolic model in which MSH from endogenously produced NO facilitates the maintenance of both NO homeostasis and signalling in S. coelicolor A3(2) M145.


Asunto(s)
Streptomyces coelicolor , Streptomyces coelicolor/genética , Streptomyces coelicolor/metabolismo , Óxido Nítrico/metabolismo , Cisteína/metabolismo , Homeostasis , Regulación Bacteriana de la Expresión Génica , Antraquinonas/metabolismo , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Antibacterianos/farmacología
3.
Biochem Biophys Res Commun ; 682: 293-298, 2023 11 19.
Artículo en Inglés | MEDLINE | ID: mdl-37832386

RESUMEN

The soybean cyst nematode (SCN) is one of the most damaging pests affecting soybean production. SCN displays important host recognition behaviors, such as hatching and infection, by recognizing several compounds produced by the host. Therefore, controlling SCN behaviors such as chemotaxis and thermotaxis is an attractive pest control strategy. In this study, we found that cyclic nucleotide-gated channels (CNG channels) regulate SCN chemotaxis and thermotaxis and Hg-tax-2, a gene encoding a CNG channel, is an important regulator of SCN behavior. Gene silencing of Hg-tax-2 and treatment with a CNG channel inhibitor reduced the attraction of second-stage juveniles to nitrate, an attractant with a different recognition mechanism from the host-derived chemoattractant(s), and to host soybean roots, as well as their avoidance behavior toward high temperatures. Co-treatment of ds Hg-tax-2 with the CNG channel inhibitor indicated that Hg-tax-2 is a major regulator of SCN chemotaxis and thermotaxis. These results suggest new avenues for research on control of SCN.


Asunto(s)
Mercurio , Nematodos , Tylenchoidea , Animales , Quimiotaxis , Canales Catiónicos Regulados por Nucleótidos Cíclicos , Glycine max/genética , Nucleótidos Cíclicos , Tylenchoidea/fisiología , Enfermedades de las Plantas
4.
J Pestic Sci ; 48(3): 107-110, 2023 Aug 20.
Artículo en Inglés | MEDLINE | ID: mdl-37745175

RESUMEN

Brassinosteroids (BRs) are steroid hormones that regulate plant growth, development, and stress resistance. In this study, we evaluated the effect of agrochemicals on dark-induced hypocotyl elongation, which is regulated by BRs, to identify novel chemicals that regulate BR action. We found that the juvenile hormone agonist fenoxycarb inhibited dark-induced hypocotyl elongation in Arabidopsis. Treatment with the same class of juvenile hormone agonist, pyriproxyfen, did not affect hypocotyl elongation. Co-treatment with fenoxycarb and BR partly canceled the fenoxycarb-induced hypocotyl suppression. In addition, gene expression analysis revealed that fenoxycarb altered the BR-responsive gene expression. These results indicate that fenoxycarb is a BR action inhibitor.

5.
FEMS Microbiol Lett ; 3702023 01 17.
Artículo en Inglés | MEDLINE | ID: mdl-37355778

RESUMEN

Our previous studies revealed that a two-component system (TCS), DevS, and DevR, regulate both nitric oxide (NO) signaling and NO homeostasis in the actinobacterium Streptomyces coelicolor A3(2) M145, suggesting a reasonable system for NO-dependent metabolism. In this study, sequence alignment of DevR and DevR homologs found Asp66 (D66) and Thr196 (T196) as predicted phosphorylation sites of DevR. Phos-tag gel electrophoretic mobility shift assay suggested that D66 and T196 are involved in the phosphorylation of DevR. The respective point mutations of D66 and T196 significantly decreased the transcriptional activity of DevR, which affected nitrite production and aerial mycelium formation. These results suggested that both D66 and T196 of DevR are important for the regulation of NO homeostasis and signaling in S. coelicolor A3(2) M145.


Asunto(s)
Streptomyces coelicolor , Fosforilación , Streptomyces coelicolor/genética , Streptomyces coelicolor/metabolismo , Óxido Nítrico/metabolismo , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Alineación de Secuencia , Regulación Bacteriana de la Expresión Génica
6.
ACS Omega ; 8(15): 13855-13862, 2023 Apr 18.
Artículo en Inglés | MEDLINE | ID: mdl-37091382

RESUMEN

Strigolactones (SLs), phytohormones that inhibit shoot branching in plants, promote the germination of root-parasitic plants, such as Striga spp. and Orobanche spp., which drastically reduces the crop yield. Therefore, reducing SL production via chemical treatment may increase the crop yield. To design specific inhibitors, it is valid to utilize the substrate structure of the target proteins as lead compounds. In this study, we focused on Os900, a rice enzyme that oxidizes the SL precursor carlactone (CL) to 4-deoxyorobanchol (4DO), and synthesized 10 CL derivatives. The effects of the synthesized CL derivatives on SL biosynthesis were evaluated by the Os900 enzyme assay in vitro and by measuring 4DO levels in rice root exudates. We identified some CL derivatives that inhibited SL biosynthesis in vitro and in vivo.

7.
Appl Environ Microbiol ; 88(23): e0122222, 2022 12 13.
Artículo en Inglés | MEDLINE | ID: mdl-36354316

RESUMEN

Nitric oxide (NO) is a well-known signaling molecule in various organisms. Streptomyces undergoes complex morphological differentiation, similar to that of fungi. A recent study revealed a nitrogen oxide metabolic cycle that forms NO in Streptomyces coelicolor A3(2) M145. Further, endogenously produced NO serves as a signaling molecule. Here, we report that endogenously produced NO regulates cyclic 3',5'-diguanylate (c-di-GMP) levels and controls aerial mycelium formation through the c-di-GMP-binding transcriptional regulator BldD in S. coelicolor A3(2) M145. These observations provide important insights into the mechanisms regulating morphological differentiation. This is the first study to demonstrate a link between NO and c-di-GMP in S. coelicolor A3(2) M145. Morphological differentiation is closely linked to the initiation of secondary metabolism in actinomycetes. Thus, the NO signaling-based regulation of aerial mycelium formation has potential applications in the fermentation industry employing useful actinomycetes. IMPORTANCE Eukaryotic and prokaryotic cells utilize nitric oxide (NO) to regulate physiological functions. Besides its role as a producer of different bioactive substances, Streptomyces is suggested to be involved in mycelial development regulated by endogenously produced NO. However, the regulatory mechanisms are unclear. In this study, we proposed that NO signaling is involved in aerial mycelium formation in S. coelicolor A3(2) M145. NO serves as a signaling molecule for the regulation of intracellular cyclic 3',5'-diguanylate (c-di-GMP) levels, resulting in aerial mycelium formation controlled by a c-di-GMP receptor, BldD. As the abundant production of valuable secondary metabolites is closely related to the initiation of morphological differentiation in Streptomyces, NO may provide value for application in industrial fermentation by serving as a tool for regulating secondary metabolism.


Asunto(s)
Streptomyces coelicolor , Streptomyces , Streptomyces coelicolor/genética , Streptomyces coelicolor/metabolismo , Óxido Nítrico/metabolismo , Regulación Bacteriana de la Expresión Génica , Proteínas Bacterianas/genética , Streptomyces/metabolismo , Micelio/metabolismo
8.
Sci Adv ; 8(44): eadd1278, 2022 11 04.
Artículo en Inglés | MEDLINE | ID: mdl-36322663

RESUMEN

Strigolactones (SLs) are a plant hormone inhibiting shoot branching/tillering and a rhizospheric, chemical signal that triggers seed germination of the noxious root parasitic plant Striga and mediates symbiosis with beneficial arbuscular mycorrhizal fungi. Identifying specific roles of canonical and noncanonical SLs, the two SL subfamilies, is important for developing Striga-resistant cereals and for engineering plant architecture. Here, we report that rice mutants lacking canonical SLs do not show the shoot phenotypes known for SL-deficient plants, exhibiting only a delay in establishing arbuscular mycorrhizal symbiosis, but release exudates with a significantly decreased Striga seed-germinating activity. Blocking the biosynthesis of canonical SLs by TIS108, a specific enzyme inhibitor, significantly lowered Striga infestation without affecting rice growth. These results indicate that canonical SLs are not the determinant of shoot architecture and pave the way for increasing crop resistance by gene editing or chemical treatment.

9.
J Pestic Sci ; 47(1): 43-46, 2022 Feb 20.
Artículo en Inglés | MEDLINE | ID: mdl-35414758

RESUMEN

Strigolactones (SLs) are carotenoid-derived plant hormones involved in several growth and developmental processes. Also, SLs are allelochemicals that induce the seed germination of root parasitic plants and the hyphal branching of arbuscular mycorrhizal fungi. In this study, to identify novel lead chemicals that inhibit SL biosynthesis, we evaluated the effect of agrochemicals on SL biosynthesis. We found that the diacylhydrazine insect growth regulator, chromafenozide, reduced the endogenous level of 4-deoxyorobanchol (4DO), a major SL in rice. Furthermore, treatment with the same class of insect growth regulator, methoxyfenozide, also resulted in the reduction of 4DO levels in rice root exudates. These results suggest that chromafenozide and methoxyfenozide are novel lead inhibitors of SL biosynthesis.

10.
Int J Mol Sci ; 22(16)2021 Aug 04.
Artículo en Inglés | MEDLINE | ID: mdl-34445089

RESUMEN

Apical periodontitis, an inflammatory lesion causing bone resorption around the apex of teeth, is treated by eradicating infectious bacteria from the root canal. However, it has a high recurrence rate and often requires retreatment. We investigated the bactericidal effect of antimicrobial photodynamic therapy (aPDT)/photodynamic antimicrobial chemotherapy (PACT) using indocyanine green (ICG)-loaded nanospheres coated with chitosan and a diode laser on a biofilm of Enterococcus faecalis, a pathogen of refractory apical periodontitis. Biofilm of E. faecalis was cultured in a porcine infected root canal model. ICG solution was injected into the root canal, which was then irradiated with a laser (810 nm wavelength) from outside the root canal. The bactericidal effect was evaluated by colony counts and scanning electron microscopy. The result of the colony counts showed a maximum 1.89 log reduction after irradiation at 2.1 W for 5 min. The temperature rise during aPDT/PACT was confirmed to be within a safe range. Furthermore, the light energy transmittance through the root was at a peak approximately 1 min after the start of irradiation, indicating that most of the ICG in the root canal was consumed. This study shows that aPDT/PACT can suppress E. faecalis in infected root canals with high efficiency.


Asunto(s)
Biopelículas/efectos de los fármacos , Enterococcus faecalis/efectos de los fármacos , Verde de Indocianina/administración & dosificación , Nanosferas , Fármacos Fotosensibilizantes/administración & dosificación , Animales , Enterococcus faecalis/fisiología , Infecciones por Bacterias Grampositivas/tratamiento farmacológico , Humanos , Verde de Indocianina/farmacología , Láseres de Semiconductores , Nanosferas/química , Fotoquimioterapia , Fármacos Fotosensibilizantes/farmacología , Porcinos
11.
Appl Environ Microbiol ; 87(14): e0048021, 2021 06 25.
Artículo en Inglés | MEDLINE | ID: mdl-33990302

RESUMEN

Nitric oxide (NO) is an important signaling molecule in eukaryotic and prokaryotic cells. A previous study revealed an NO synthase-independent NO production metabolic cycle in which the three nitrogen oxides, nitrate (NO3-), nitrite (NO2-), and NO, were generated in the actinobacterium Streptomyces coelicolor A3(2). NO was suggested to act as a signaling molecule, functioning as a hormone that regulates secondary metabolism. Here, we demonstrate the NO-mediated regulation of the production of the blue-pigmented antibiotic actinorhodin (ACT), via the heme-based DevS/R two-component system (TCS). Intracellular NO controls the stabilization or inactivation of DevS, depending on the NO concentration. An electrophoretic mobility shift assay and chromatin immunoprecipitation-quantitative PCR analysis revealed the direct binding between DevR and the promoter region of actII-ORF4, resulting in gene expression. Our results indicate that NO regulates the DevS/R TCS, thereby strictly controlling the secondary metabolism of S. coelicolor A3(2). IMPORTANCE Diverse organisms, such as mammals, plants, and bacteria, utilize NO via well-known signal transduction mechanisms. Many useful secondary metabolite-producing bacteria of the Streptomyces genus had been also suggested for the metabolism regulated by endogenously produced NO; however, the regulatory mechanisms remain to be elucidated. In this study, we demonstrated the molecular mechanism by which endogenously produced NO regulates antibiotic production via the DevS/R TCS in S. coelicolor A3(2). NO serves as both a stabilizer and a repressor in the regulation of antibiotic production. This report shows the mechanism by which Streptomyces utilizes endogenously produced NO to modulate its normal life cycle. Moreover, this study implies that studying NO signaling in actinobacteria can help in the development of both clinical strategies against pathogenic actinomycetes and the actinobacterial industries.


Asunto(s)
Óxido Nítrico/metabolismo , Streptomyces coelicolor/metabolismo , Actinas/genética , Antraquinonas/metabolismo , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Regulación Bacteriana de la Expresión Génica , Proteínas del Helminto/genética , Regiones Promotoras Genéticas , Metabolismo Secundario , Streptomyces coelicolor/genética , Factores de Transcripción/genética , Factores de Transcripción/metabolismo
12.
Biochim Biophys Acta Proteins Proteom ; 1869(7): 140644, 2021 07.
Artículo en Inglés | MEDLINE | ID: mdl-33716191

RESUMEN

Microbacterium hydrocarbonoxydans has been isolated using an unnatural acylhydrazide compound as the sole carbon source. The compound is hydrolyzed by bacterial hydrazidase, and the gene expression of the enzyme is considered to be controlled by a transcription factor of the Isocitrate lyase Regulator (IclR) family, belonging to the one-component signaling systems. Recently, we reported the crystal structure of an unliganded IclR homolog from M. hydrocarbonoxydans, named putative 4-hydroxybenzoate response regulator (pHbrR), which has a unique homotetramer conformation. In this study, we report the crystal structure of pHbrR complexed with 4-hydroxybenzoic acid, the catalytic product of hydrazidase, at 2.0 Å resolution. pHbrR forms a homodimer with multimeric rearrangement in the unliganded state. Gel filtration column chromatography results suggested dimer-tetramer rearrangement. We observed conformational change in the loop region covering the ligand-binding site, and domain rearrangements in the monomer. This study reports the first liganded IclR family protein structure that demonstrates large structural rearrangements between liganded and unliganded proteins, which may represent a general model for IclRs.


Asunto(s)
Isocitratoliasa/metabolismo , Factores de Transcripción/metabolismo , Proteínas Bacterianas/química , Sitios de Unión , Cristalografía por Rayos X/métodos , Proteínas de Escherichia coli/metabolismo , Proteínas de Escherichia coli/ultraestructura , Isocitratoliasa/ultraestructura , Isocitratos , Ligandos , Microbacterium/metabolismo , Modelos Moleculares , Conformación Proteica , Proteínas Represoras/metabolismo , Proteínas Represoras/ultraestructura , Factores de Transcripción/ultraestructura
13.
Molecules ; 25(23)2020 Nov 25.
Artículo en Inglés | MEDLINE | ID: mdl-33255720

RESUMEN

Strigolactones (SLs) are carotenoid-derived plant hormones involved in the development of various plants. SLs also stimulate seed germination of the root parasitic plants, Striga spp. and Orobanche spp., which reduce crop yield. Therefore, regulating SL biosynthesis may lessen the damage of root parasitic plants. Biosynthetic inhibitors effectively control biological processes by targeted regulation of biologically active compounds. In addition, biosynthetic inhibitors regulate endogenous levels in developmental stage- and tissue-specific manners. To date, although some chemicals have been found as SL biosynthesis inhibitor, these are derived from only three lead chemicals. In this study, to find a novel lead chemical for SL biosynthesis inhibitor, 27 nitrogen-containing heterocyclic derivatives were screened for inhibition of SL biosynthesis. Triflumizole most effectively reduced the levels of rice SL, 4-deoxyorobanchol (4DO), in root exudates. In addition, triflumizole inhibited endogenous 4DO biosynthesis in rice roots by inhibiting the enzymatic activity of Os900, a rice enzyme that converts the SL intermediate carlactone to 4DO. A Striga germination assay revealed that triflumizole-treated rice displayed a reduced level of germination stimulation for Striga. These results identify triflumizole as a novel lead compound for inhibition of SL biosynthesis.


Asunto(s)
Vías Biosintéticas/efectos de los fármacos , Compuestos Heterocíclicos con 3 Anillos/metabolismo , Imidazoles/farmacología , Lactonas/metabolismo , Germinación/efectos de los fármacos , Imidazoles/química , Estructura Molecular , Oryza/efectos de los fármacos , Oryza/metabolismo , Raíces de Plantas/efectos de los fármacos
14.
Biochem Biophys Res Commun ; 525(3): 720-725, 2020 05 07.
Artículo en Inglés | MEDLINE | ID: mdl-32143826

RESUMEN

Microbacterium hydrocarbonoxydans was isolated, using hydrazide compounds as its sole carbon source. The key enzyme that metabolizes these compounds was identified as hydrazidase, and the operon containing the gene coding for the enzyme, was revealed by genome sequencing. The operon also contained genes coding for an ATP-binding cassette transporter (ABC transporter), which was expected to transport the hydrazide compounds. Substrate binding protein (SBP), a component subunit of the transporter, plays an important role in recognizing the correct substrates for transport. Therefore, to elucidate the mechanism of recognition of the unnatural hydrazide compounds, we determined the crystal structures of the SBP, obtained from M. hydrocarbonoxydans (Mh-SBP), complexed with and without the hydrazide compound, at 2.2 Å and 1.75 Å resolutions, respectively. The overall structures of Mh-SBP were similar to those of the SBP in oligopeptide transporters such as OppA. On comparison, the liganded and unliganded structures of Mh-SBP showed an open - close conformation change. Interestingly, the binding mode of the compound to Mh-SBP was almost identical to that of the compound to hydrazidase, suggesting that the ABC transporter served transporting these compounds. Furthermore, based on the hydrazide complex structure, paraben, the other putative substrate of the protein, was successfully used with Mh-SBP to obtain the paraben complex structure.


Asunto(s)
Actinobacteria/metabolismo , Proteínas Bacterianas/química , Proteínas Bacterianas/metabolismo , Hidrazinas/metabolismo , Proteínas de Transporte de Membrana/química , Proteínas de Transporte de Membrana/metabolismo , Ligandos , Microbacterium , Modelos Moleculares , Parabenos/química , Parabenos/metabolismo , Relación Estructura-Actividad , Especificidad por Sustrato
15.
Biosci Biotechnol Biochem ; 84(4): 734-742, 2020 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-31842701

RESUMEN

scyllo-inositol dehydrogenase, isolated from Paracoccus laeviglucosivorans (Pl-sIDH), exhibits a broad substrate specificity: it oxidizes scyllo- and myo-inositols as well as L-glucose, converting L-glucose to L-glucono-1,5-lactone. Based on the crystal structures previously reported, Arg178 residue, located at the entry port of the catalytic site, seemed to be important for accepting substrates. Here, we report the role of Arg178 by using an alanine-substituted mutant for kinetic analysis as well as to determine the crystal structures. The wild-type Pl-sIDH exhibits the activity for scyllo-inositol most preferably followed by myo-inositol and L-glucose. On the contrary, the R178A mutant abolished the activities for both inositols, but remained active for L-glucose to the same extent as its wild-type. Based on the crystal structures of the mutant, the side chain of Asp191 flipped out of the substrate binding site. Therefore, Arg178 is important in positioning Asp191 correctly to exert its catalytic activities.Abbreviations: IDH: inositol dehydrogenase; LB: Luria-Bertani; kcat: catalyst rate constant; Km: Michaelis constant; NAD: nicotinamide dinucleotide; NADH: nicotinamide dinucleotide reduced form; PDB; Protein Data Bank; PDB entry: 6KTJ, 6KTK, 6KTL.


Asunto(s)
Sustitución de Aminoácidos , Glucosa/metabolismo , Inositol/metabolismo , Oxidorreductasas/metabolismo , Paracoccus/enzimología , Cinética , Oxidorreductasas/aislamiento & purificación , Conformación Proteica , Especificidad por Sustrato
16.
Acta Crystallogr F Struct Biol Commun ; 75(Pt 9): 599-607, 2019 Sep 01.
Artículo en Inglés | MEDLINE | ID: mdl-31475927

RESUMEN

Hygromycin B (HygB) is one of the aminoglycoside antibiotics, and it is widely used as a reagent in molecular-biology experiments. Two kinases are known to inactivate HygB through phosphorylation: aminoglycoside 7''-phosphotransferase-Ia [APH(7'')-Ia] from Streptomyces hygroscopicus and aminoglycoside 4-phosphotransferase-Ia [APH(4)-Ia] from Escherichia coli. They phosphorylate the hydroxyl groups at positions 7'' and 4 of the HygB molecule, respectively. Previously, the crystal structure of APH(4)-Ia was reported as a ternary complex with HygB and 5'-adenylyl-ß,γ-imidodiphosphate (AMP-PNP). To investigate the differences in the substrate-recognition mechanism between APH(7'')-Ia and APH(4)-Ia, the crystal structure of APH(7'')-Ia complexed with HygB is reported. The overall structure of APH(7'')-Ia is similar to those of other aminoglycoside phosphotransferases, including APH(4)-Ia, and consists of an N-terminal lobe (N-lobe) and a C-terminal lobe (C-lobe). The latter also comprises a core and a helical domain. Accordingly, the APH(7'')-Ia and APH(4)-Ia structures fit globally when the structures are superposed at three catalytically important conserved residues, His, Asp and Asn, in the Brenner motif, which is conserved in aminoglycoside phosphotransferases as well as in eukaryotic protein kinases. On the other hand, the phosphorylated hydroxyl groups of HygB in both structures come close to the Asp residue, and the HygB molecules in each structure lie in opposite directions. These molecules were held by the helical domain in the C-lobe, which exhibited structural differences between the two kinases. Furthermore, based on the crystal structures of APH(7'')-Ia and APH(4)-Ia, some mutated residues in their thermostable mutants reported previously were located at the same positions in the two enzymes.


Asunto(s)
Antibacterianos/química , Higromicina B/química , Kanamicina Quinasa/química , Streptomyces/enzimología , Adenilil Imidodifosfato/química , Secuencias de Aminoácidos/genética , Aminoglicósidos/química , Sitios de Unión , Catálisis , Cristalografía por Rayos X , Escherichia coli/metabolismo , Kanamicina Quinasa/genética , Kanamicina Quinasa/metabolismo , Fosforilación , Dominios Proteicos , Especificidad por Sustrato
17.
J Agric Food Chem ; 67(22): 6143-6149, 2019 Jun 05.
Artículo en Inglés | MEDLINE | ID: mdl-31083983

RESUMEN

Strigolactones (SLs) are one of the plant hormones that control several important agronomic traits, such as shoot branching, leaf senescence, and stress tolerance. Manipulation of the SL biosynthesis can increase the crop yield. We previously reported that a triazole derivative, TIS108, inhibits SL biosynthesis. In this study, we synthesized a number of novel TIS108 derivatives. Structure-activity relationship studies revealed that 4-(2-phenoxyethoxy)-1-phenyl-2-(1 H-1,2,4-triazol-1-yl)butan-1-one (KK5) inhibits the level of 4-deoxyorobanchol in roots more strongly than TIS108. We further found that KK5-treated Arabidopsis showed increased branching phenotype with the upregulated gene expression of AtMAX3 and AtMAX4. These results indicate that KK5 is a specific SL biosynthesis inhibitor in rice and Arabidopsis.


Asunto(s)
Reguladores del Crecimiento de las Plantas/antagonistas & inhibidores , Terpenos/antagonistas & inhibidores , Triazoles/química , Triazoles/farmacología , Arabidopsis/efectos de los fármacos , Arabidopsis/genética , Arabidopsis/metabolismo , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Oryza/efectos de los fármacos , Oryza/genética , Oryza/metabolismo , Reguladores del Crecimiento de las Plantas/biosíntesis , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Terpenos/metabolismo , Triazoles/síntesis química
18.
Ann Thorac Cardiovasc Surg ; 25(1): 32-38, 2019 Feb 20.
Artículo en Inglés | MEDLINE | ID: mdl-30122739

RESUMEN

OBJECTIVE: The aim of this retrospective study was to investigate the early operative results and detect the factors influencing the fate of radial artery grafts (RAGs) by evaluating the mid-term patency. METHODS: We retrospectively reviewed 410 patients who underwent isolated coronary artery bypass grafting using RAG. RAGs were anastomosed to 526 coronary arteries. Mid-term angiography was performed in 214 patients at an average 4.9 years after the operation. RESULTS: The early patency of RAGs was 97.6%. Cumulative 5-year patency was 86.5% for RAG, 94.1% for LITA graft, and 81.0% for saphenous vein graft (SVG). RAG was significantly superior to SVG in mid-term patency. Individual grafting (not sequential grafting) (hazard ratio [HR]: 2.535; 95% confidence interval [CI]: 1.293-5.281; p = 0.006) and grafting to the target coronary artery with ≤75% proximal stenosis (HR: 1.947; 95% CI: 1.090-3.484; p = 0.025) were found to be independent risk factors influencing late RAG patency. CONCLUSIONS: The patency of RAGs was superior to that of SVGs in the studied population. When using RAGs, grafting to the target vessel with severe proximal stenosis is favorable. The RAG is suitable for sequential grafting.


Asunto(s)
Puente de Arteria Coronaria/efectos adversos , Puente de Arteria Coronaria/métodos , Estenosis Coronaria/cirugía , Oclusión de Injerto Vascular/etiología , Arteria Radial/trasplante , Anciano , Angiografía por Tomografía Computarizada , Angiografía Coronaria , Estenosis Coronaria/diagnóstico por imagen , Estenosis Coronaria/fisiopatología , Femenino , Oclusión de Injerto Vascular/diagnóstico por imagen , Oclusión de Injerto Vascular/fisiopatología , Humanos , Masculino , Persona de Mediana Edad , Tomografía Computarizada Multidetector , Arteria Radial/diagnóstico por imagen , Arteria Radial/fisiopatología , Estudios Retrospectivos , Factores de Riesgo , Vena Safena/diagnóstico por imagen , Vena Safena/fisiopatología , Vena Safena/trasplante , Índice de Severidad de la Enfermedad , Factores de Tiempo , Insuficiencia del Tratamiento , Grado de Desobstrucción Vascular
19.
Kyobu Geka ; 71(13): 1081-1083, 2018 12.
Artículo en Japonés | MEDLINE | ID: mdl-30587746

RESUMEN

A unicuspid aortic valve is an extremely rare congenital aortic valvular abnormality. We herein present 2 cases of unicuspid aortic valve diagnosed based on intraoperative findings. In case 1, a 75-year-old man was admitted to our hospital because of severe aortic regurgitation. We performed aortic valve replacement using a bioprosthetic valve, and a unicuspid aortic valve was definitively diagnosed according to the intraoperative findings. In case 2, a 54-year-old man developed dyspnea due to severe aortic stenosis. Aortic valve replacement using mechanical valve was performed, and we were able to diagnose unicuspid aortic valve intraoperatively. Achieving a preoperative definitive diagnosis of congenital unicuspid aortic valve by transthoracic echocardiography is reportedly difficult;however, transesophageal echocardiography may be effective for preoperative definitive diagnosis.


Asunto(s)
Insuficiencia de la Válvula Aórtica/cirugía , Estenosis de la Válvula Aórtica/cirugía , Válvula Aórtica/anomalías , Implantación de Prótesis de Válvulas Cardíacas , Prótesis Valvulares Cardíacas , Anciano , Insuficiencia de la Válvula Aórtica/complicaciones , Estenosis de la Válvula Aórtica/complicaciones , Bioprótesis , Ecocardiografía , Humanos , Masculino , Persona de Mediana Edad
20.
PLoS One ; 13(5): e0198010, 2018.
Artículo en Inglés | MEDLINE | ID: mdl-29799855

RESUMEN

For about 70 years, L-glucose had been considered non-metabolizable by either mammalian or bacterial cells. Recently, however, an L-glucose catabolic pathway has been discovered in Paracoccus laeviglucosivorans, and the genes responsible cloned. Scyllo-inositol dehydrogenase is involved in the first step in the pathway that oxidizes L-glucose to produce L-glucono-1,5-lactone with concomitant reduction of NAD+ dependent manner. Here, we report the crystal structure of the ternary complex of scyllo-inositol dehydrogenase with NAD+ and L-glucono-1,5-lactone at 1.8 Å resolution. The enzyme adopts a homo-tetrameric structure, similar to those of the inositol dehydrogenase family, and the electron densities of the bound sugar was clearly observed, allowing identification of the residues responsible for interaction with the substrate in the catalytic site. In addition to the conserved catalytic residues (Lys106, Asp191, and His195), another residue, His318, located in the loop region of the adjacent subunit, is involved in substrate recognition. Site-directed mutagenesis confirmed the role of these residues in catalytic activity. We also report the complex structures of the enzyme with myo-inositol and scyllo-inosose. The Arg178 residue located in the flexible loop at the entrance of the catalytic site is also involved in substrate recognition, and plays an important role in accepting both L-glucose and inositols as substrates. On the basis of these structural features, which have not been identified in the known inositol dehydrogenases, and a phylogenetic analysis of IDH family enzymes, we suggest a novel subfamily of the GFO/IDH/MocA family. Since many enzymes in this family have not biochemically characterized, our results could promote to find their activities with various substrates.


Asunto(s)
Glucosa/metabolismo , Inositol/metabolismo , Deshidrogenasas del Alcohol de Azúcar/química , Deshidrogenasas del Alcohol de Azúcar/metabolismo , Secuencia de Aminoácidos , Modelos Moleculares , Mutación , Oxidación-Reducción , Conformación Proteica , Deshidrogenasas del Alcohol de Azúcar/genética
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