Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 915
Filtrar
1.
Elife ; 112022 12 08.
Artigo em Inglês | MEDLINE | ID: mdl-36476508

RESUMO

We aimed to elucidate the evolutionary trajectories of gallbladder adenocarcinoma (GBAC) using multi-regional and longitudinal tumor samples. Using whole-exome sequencing data, we constructed phylogenetic trees in each patient and analyzed mutational signatures. A total of 11 patients including 2 rapid autopsy cases were enrolled. The most frequently altered gene in primary tumors was ERBB2 and TP53 (54.5%), followed by FBXW7 (27.3%). Most mutations in frequently altered genes in primary tumors were detectable in concurrent precancerous lesions (biliary intraepithelial neoplasia [BilIN]), but a substantial proportion was subclonal. Subclonal diversity was common in BilIN (n=4). However, among subclones in BilIN, a certain subclone commonly shrank in concurrent primary tumors. In addition, selected subclones underwent linear and branching evolution, maintaining subclonal diversity. Combined analysis with metastatic tumors (n=11) identified branching evolution in nine patients (81.8%). Of these, eight patients (88.9%) had a total of 11 subclones expanded at least sevenfold during metastasis. These subclones harbored putative metastasis-driving mutations in cancer-related genes such as SMAD4, ROBO1, and DICER1. In mutational signature analysis, six mutational signatures were identified: 1, 3, 7, 13, 22, and 24 (cosine similarity >0.9). Signatures 1 (age) and 13 (APOBEC) decreased during metastasis while signatures 22 (aristolochic acid) and 24 (aflatoxin) were relatively highlighted. Subclonal diversity arose early in precancerous lesions and clonal selection was a common event during malignant transformation in GBAC. However, selected cancer clones continued to evolve and thus maintained subclonal diversity in metastatic tumors.


Assuntos
Adenocarcinoma , Lesões Pré-Cancerosas , Humanos , Adolescente , Filogenia , Vesícula Biliar , Proteínas do Tecido Nervoso , Receptores Imunológicos , Adenocarcinoma/genética , Mutação , Lesões Pré-Cancerosas/genética , Pigmentos Biliares , Ribonuclease III , RNA Helicases DEAD-box
2.
Eur J Med Res ; 27(1): 224, 2022 Oct 29.
Artigo em Inglês | MEDLINE | ID: mdl-36309733

RESUMO

Bile pigments, such as bilirubin and biliverdin, are end products of the heme degradation pathway in mammals and are widely known for their cytotoxic effects. However, recent studies have revealed that they exert cytoprotective effects through antioxidative, anti-inflammatory, and immunosuppressive properties. All these mechanisms are indispensable in the treatment of diseases in the field of emergency and critical care medicine, such as coronary ischemia, stroke, encephalomyelitis, acute lung injury/acute respiratory distress syndrome, mesenteric ischemia, and sepsis. While further research is required before the safe application of bile pigments in the clinical setting, their underlying mechanisms shed light on their utilization as therapeutic agents in the field of emergency and critical care medicine. This article aims to summarize the current understanding of bile pigments and re-evaluate their therapeutic potential in the diseases listed above.


Assuntos
Pigmentos Biliares , Síndrome do Desconforto Respiratório , Animais , Humanos , Pigmentos Biliares/metabolismo , Biliverdina/metabolismo , Antioxidantes/uso terapêutico , Cuidados Críticos , Mamíferos/metabolismo
3.
J Agric Food Chem ; 70(38): 12055-12064, 2022 Sep 28.
Artigo em Inglês | MEDLINE | ID: mdl-36122349

RESUMO

Cannabidiol (CBD), the main nonpsychoactive cannabinoid in Cannabis sativa, has diverse applications in the pharmacological, food, and cosmetic industries. The long plantation period and the complex chemical structure of cannabidiol pose a great challenge on CBD supply. Here, we achieved de novo biosynthesis of cannabidiol in Saccharomyces cerevisiae. The CBD production was further enhanced by 2.53-fold through pushing the supply of precursors and fusion protein construction. Bile pigment transporter 1 (BPT1) was the most effective transporter for transferring cannabigerolic acid (CBGA) from the cytoplasm to the vacuole, which removed the physical barrier separating CBGA and its catalytic enzyme. The lowest binding energy of the CBGA-BPT1 complex confirmed a strong interaction between BPT1 and CBGA. A CBD yield of 6.92 mg/L was achieved, which was 100-fold higher than the yield generated by the starting strain. This study provides insights into high-level CBD-producing strain construction and lays the foundation for CBD supply.


Assuntos
Canabidiol , Canabinoides , Cannabis , Pigmentos Biliares , Canabidiol/química , Cannabis/química , Cannabis/genética , Saccharomyces cerevisiae/genética , Vacúolos
4.
Br J Cancer ; 127(9): 1603-1614, 2022 11.
Artigo em Inglês | MEDLINE | ID: mdl-36068277

RESUMO

BACKGROUND: Intraductal papillary neoplasms (IPN) and biliary epithelial neoplasia (BilIN) are well-defined precursor lesions of biliary tract carcinoma (BTC). The aim of this study was to provide a comprehensive characterisation of the inflammatory microenvironment in BTC precursor lesions. METHODS: Immunohistochemistry was employed to assess tumour-infiltrating immune cells in tissue samples from patients, for whom precursor lesions were identified alongside invasive BTC. The spatiotemporal evolution of the immune microenvironment during IPN-associated carcinogenesis was comprehensively analysed using triplet sample sets of non-neoplastic epithelium, precursor lesion and invasive BTC. Immune-cell dynamics during IPN- and BilIN-associated carcinogenesis were subsequently compared. RESULTS: Stromal CD3+ (P = 0.002), CD4+ (P = 0.007) and CD8+ (P < 0.001) T cells, CD20+ B cells (P = 0.008), MUM1+ plasma cells (P = 0.012) and CD163+ M2-like macrophages (P = 0.008) significantly decreased in IPN compared to non-tumorous biliary epithelium. Upon transition from IPN to invasive BTC, stromal CD68+ (P = 0.001) and CD163+ (P < 0.001) macrophages significantly increased. In contrast, BilIN-driven carcinogenesis was characterised by significant reduction of intraepithelial CD8+ T-lymphocytic infiltration from non-tumorous epithelium via BilIN (P = 0.008) to BTC (P = 0.004). CONCLUSION: IPN and BilIN are immunologically distinct entities that undergo different immune-cell variations during biliary carcinogenesis. Intraepithelial CD8+ T-lymphocytic infiltration of biliary tissue decreased already at the IPN-precursor stage, whereas BilIN-associated carcinogenesis showed a slowly progressing reduction towards invasive carcinoma.


Assuntos
Neoplasias dos Ductos Biliares , Neoplasias do Sistema Biliar , Sistema Biliar , Colangiocarcinoma , Humanos , Colangiocarcinoma/patologia , Neoplasias dos Ductos Biliares/patologia , Sistema Biliar/patologia , Neoplasias do Sistema Biliar/patologia , Carcinogênese/patologia , Ductos Biliares Intra-Hepáticos/patologia , Análise Espaço-Temporal , Pigmentos Biliares , Microambiente Tumoral
5.
J Proteome Res ; 21(10): 2261-2276, 2022 Oct 07.
Artigo em Inglês | MEDLINE | ID: mdl-36169658

RESUMO

Malaria varies in severity, with complications ranging from uncomplicated to severe malaria. Severe malaria could be attributed to peripheral hyperparasitemia or cerebral malaria. The metabolic interactions between the host and Plasmodium species are yet to be understood during these infections of varied pathology and severity. An untargeted metabolomics approach utilizing the liquid chromatography-mass spectrometry platform has been used to identify the affected host metabolic pathways and associated metabolites in the serum of murine malaria models with uncomplicated malaria, hyperparasitemia, and experimental cerebral malaria. We report that mice with malaria share similar metabolic attributes like higher levels of bile acids, bile pigments, and steroid hormones that have been reported for human malaria infections. Moreover, in severe malaria, upregulated levels of metabolites like phenylalanine, histidine, valine, pipecolate, ornithine, and pantothenate, with decreased levels of arginine and hippurate, were observed. Metabolites of sphingolipid metabolism were upregulated in experimental cerebral malaria. Higher levels of 20-hydroxy-leukotriene B4 and epoxyoctadecamonoenoic acids were found in uncomplicated malaria, with lower levels observed for experimental cerebral malaria. Our study provides insights into host biology during different pathological stages of malaria disease and would be useful for the selection of animal models for evaluating diagnostic and therapeutic interventions against malaria. The raw data files are available via MetaboLights with the identifier MTBLS4387.


Assuntos
Malária Cerebral , Animais , Arginina , Ácidos e Sais Biliares , Pigmentos Biliares , Modelos Animais de Doenças , Hipuratos , Histidina , Hormônios , Humanos , Camundongos , Ornitina , Fenilalanina , Plasmodium berghei , Esfingolipídeos , Valina
6.
Biochemistry ; 61(14): 1444-1455, 2022 07 19.
Artigo em Inglês | MEDLINE | ID: mdl-35759789

RESUMO

Cyanobacteriochromes (CBCRs) are photoreceptors consisting of single or tandem GAF (cGMP-phosphodiesterase/adenylate cyclase/FhlA) domains that bind bilin chromophores. Canonical red/green CBCR GAF domains are a well-characterized subgroup of the expanded red/green CBCR GAF domain family that binds phycocyanobilin (PCB) and converts between a thermally stable red-absorbing Pr state and a green-absorbing Pg state. The rate of thermal reversion from Pg to Pr varies widely among canonical red/green CBCR GAF domains, with half-lives ranging from days to seconds. Since the thermal reversion rate is an important parameter for the application of CBCR GAF domains as optogenetic tools, the molecular factors controlling the thermal reversion rate are of particular interest. Here, we report that point mutations in a well-conserved W(S/G)GE motif alter reversion rates in canonical red/green CBCR GAF domains in a predictable manner. Specifically, S-to-G mutations enhance thermal reversion rates, while the reverse, G-to-S mutations slow thermal reversion. Despite the distance (>10 Å) of the mutation site from the chromophore, molecular dynamics simulations and nuclear magnetic resonance (NMR) analyses suggest that the presence of a glycine residue allows the formation of a water bridge that alters the conformational dynamics of chromophore-interacting residues, leading to enhanced Pg to Pr thermal reversion.


Assuntos
Fotorreceptores Microbianos , Fitocromo , Adenilil Ciclases/metabolismo , Proteínas de Bactérias/química , Pigmentos Biliares , Luz , Conformação Molecular , Mutação , Fotorreceptores Microbianos/química , Fitocromo/química
7.
Photochem Photobiol Sci ; 21(4): 447-469, 2022 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-35394641

RESUMO

Cyanobacteriochromes (CBCRs) are phytochrome-related photosensory proteins that play an essential role in regulating phototaxis, chromatic acclimation, and cell aggregation in cyanobacteria. Here, we apply solid-state NMR spectroscopy to the red/green GAF2 domain of the CBCR AnPixJ assembled in vitro with a uniformly 13C- and 15N-labeled bilin chromophore, tracking changes in electronic structure, geometry, and structural heterogeneity of the chromophore as well as intimate contacts between the chromophore and protein residues in the photocycle. Our data confirm that the bilin ring D is strongly twisted with respect to the B-C plane in both dark and photoproduct states. We also identify a greater structural heterogeneity of the bilin chromophore in the photoproduct than in the dark state. In addition, the binding pocket is more hydrated in the photoproduct. Observation of interfacial 1H contacts of the photoproduct chromophore, together with quantum mechanics/molecular mechanics (QM/MM)-based structural models for this photoproduct, clearly suggests the presence of a biprotonated (cationic) imidazolium side-chain for a conserved histidine residue (322) at a distance of ~2.7 Å, generalizing the recent theoretical findings that explicitly link the structural heterogeneity of the dark-state chromophore to the protonation of this specific residue. Moreover, we examine pH effects on this in vitro assembled holoprotein, showing a substantially altered electronic structure and protonation of the photoproduct chromophore even with a small pH drop from 7.8 to 7.2. Our studies provide further information regarding the light- and pH-induced changes of the chromophore and the rearrangements of the hydrogen-bonding and electrostatic interaction network around it. Possible correlations between structural heterogeneity of the chromophore, protonation of the histidine residue nearby, and hydration of the pocket in both photostates are discussed.


Assuntos
Fotorreceptores Microbianos , Fitocromo , Proteínas de Bactérias/química , Pigmentos Biliares/química , Pigmentos Biliares/metabolismo , Histidina , Concentração de Íons de Hidrogênio , Luz , Fotorreceptores Microbianos/química , Fitocromo/metabolismo
8.
Structure ; 30(4): 534-536, 2022 04 07.
Artigo em Inglês | MEDLINE | ID: mdl-35395194

RESUMO

Attachment of bilins to phycobiliproteins is performed by dedicated lyases. In this issue of Structure, Kumarapperuma et al., 2022 present the structure of an E/F type lyase-isomerase that identifies the correct biological interface between active domains, suggesting that a previous E/F lyase misidentified the heterodimer structure from the crystal lattice.


Assuntos
Liases , Pigmentos Biliares , Liases/química , Ficobiliproteínas/química
9.
J Phys Chem B ; 126(14): 2647-2657, 2022 04 14.
Artigo em Inglês | MEDLINE | ID: mdl-35357137

RESUMO

The ability of phytochromes to act as photoswitches in plants and microorganisms depends on interactions between a bilin-like chromophore and a host protein. The interconversion occurs between the spectrally distinct red (Pr) and far-red (Pfr) conformers. This conformational change is triggered by the photoisomerization of the chromophore D-ring pyrrole. In this study, as a representative example of a phytochrome-bilin system, we consider biliverdin IXα (BV) bound to bacteriophytochrome (BphP) from Deinococcus radiodurans. In the absence of light, we use an enhanced sampling molecular dynamics (MD) method to overcome the photoisomerization energy barrier. We find that the calculated free energy (FE) barriers between essential metastable states agree with spectroscopic results. We show that the enhanced dynamics of the BV chromophore in BphP contributes to triggering nanometer-scale conformational movements that propagate by two experimentally determined signal transduction pathways. Most importantly, we describe how the metastable states enable a thermal transition known as the dark reversion between Pfr and Pr, through a previously unknown intermediate state of Pfr. We present the heterogeneity of temperature-dependent Pfr states at the atomistic level. This work paves a way toward understanding the complete mechanism of the photoisomerization of a bilin-like chromophore in phytochromes.


Assuntos
Fitocromo , Proteínas de Bactérias/química , Pigmentos Biliares , Biliverdina/química , Sítios de Ligação , Conformação Molecular , Fitocromo/química
10.
Cancer Res ; 82(9): 1803-1817, 2022 05 03.
Artigo em Inglês | MEDLINE | ID: mdl-35247892

RESUMO

Biliary cancer has long been known to carry a poor prognosis, yet the molecular pathogenesis of carcinoma of the extrahepatic biliary system and its precursor lesions remains elusive. Here we investigated the role of Kras and canonical Wnt pathways in the tumorigenesis of the extrahepatic bile duct (EHBD) and gall bladder (GB). In mice, concurrent activation of Kras and Wnt pathways induced biliary neoplasms that resembled human intracholecystic papillary-tubular neoplasm (ICPN) and biliary intraepithelial neoplasia (BilIN), putative precursors to invasive biliary cancer. At a low frequency, these lesions progressed to adenocarcinoma in a xenograft model, establishing them as precancerous lesions. Global gene expression analysis revealed increased expression of genes associated with c-Myc and TGFß pathways in mutant biliary spheroids. Silencing or pharmacologic inhibition of c-Myc suppressed proliferation of mutant biliary spheroids, whereas silencing of Smad4/Tgfbr2 or pharmacologic inhibition of TGFß signaling increased proliferation of mutant biliary spheroids and cancer formation in vivo. Human ICPNs displayed activated Kras and Wnt signals and c-Myc and TGFß pathways. Thus, these data provide direct evidence that concurrent activation of the Kras and canonical Wnt pathways results in formation of ICPN and BilIN, which could develop into biliary cancer. SIGNIFICANCE: This work shows how dysregulation of canonical cell growth pathways drives precursors to biliary cancers and identifies several molecular vulnerabilities as potential therapeutic targets in these precursors to prevent oncogenic progression.


Assuntos
Neoplasias dos Ductos Biliares , Neoplasias do Sistema Biliar , Carcinoma in Situ , Lesões Pré-Cancerosas , Animais , Neoplasias dos Ductos Biliares/genética , Neoplasias dos Ductos Biliares/patologia , Pigmentos Biliares/metabolismo , Neoplasias do Sistema Biliar/genética , Carcinoma in Situ/patologia , Humanos , Camundongos , Lesões Pré-Cancerosas/patologia , Proteínas Proto-Oncogênicas p21(ras)/genética , Proteínas Proto-Oncogênicas p21(ras)/metabolismo , Fator de Crescimento Transformador beta/metabolismo , Via de Sinalização Wnt/genética
11.
Small Methods ; 6(4): e2101359, 2022 04.
Artigo em Inglês | MEDLINE | ID: mdl-35142112

RESUMO

Endogenic pigments derived from hemoglobin have been successfully applied in the clinic for both imaging and therapy based on their inherent photophysical and photochemical properties, including light absorption, fluorescence emission, and producing reactive oxygen species. However, the clinically approved endogenic pigments can be excited only by UV/vis light, restricting the penetration depth of in vivo applications. Recently, endogenic pigments with NIR-absorbing properties have been explored for constructing functional nanomaterials. Here, the overview of NIR-absorbing endogenic pigments, mainly bile pigments, and melanins, as emerging building blocks for supramolecular construction of diagnostic and therapeutic nanomaterials is provided. The endogenic origins, synthetic pathways, and structural characteristics of the NIR-absorbing endogenic pigments are described. The self-assembling approaches and noncovalent interactions in fabricating the nanomaterials are emphasized. Since bile pigments and melanins are inherently photothermal agents, the resulting nanomaterials are demonstrated as promising candidates for photoacoustic imaging and photothermal therapy. Integration of additional diagnostic and therapeutic agents by the nanomaterials through chemical conjugation or physical encapsulation toward synergetic effects is also included. Especially, the degradation behaviors of the nanomaterials in biological environments are summarized. Along with the challenges, future perspectives are discussed for accelerating the ration design and clinical translation of NIR-absorbing nanomaterials.


Assuntos
Melaninas , Nanoestruturas , Pigmentos Biliares , Nanoestruturas/uso terapêutico , Fototerapia , Nanomedicina Teranóstica/métodos
12.
Structure ; 30(4): 564-574.e3, 2022 04 07.
Artigo em Inglês | MEDLINE | ID: mdl-35148828

RESUMO

Chromophore attachment of the light-harvesting apparatus represents one of the most important post-translational modifications in photosynthetic cyanobacteria. Extensive pigment diversity of cyanobacteria critically depends on bilin lyases that covalently attach chemically distinct chromophores to phycobiliproteins. However, how bilin lyases catalyze bilin ligation reactions and how some lyases acquire additional isomerase abilities remain elusive at the molecular level. Here, we report the crystal structure of a representative bilin lyase-isomerase MpeQ. This structure has revealed a "question-mark" protein architecture that unambiguously establishes the active site conserved among the E/F-type bilin lyases. Based on structural, mutational, and modeling data, we demonstrate that stereoselectivity of the active site plays a critical role in conferring the isomerase activity of MpeQ. We further advance a tyrosine-mediated reaction scheme unifying different types of bilin lyases. These results suggest that lyases and isomerase actions of bilin lyases arise from two coupled molecular events of distinct origin.


Assuntos
Cianobactérias , Liases , Pigmentos Biliares/metabolismo , Cianobactérias/metabolismo , Isomerases/genética , Isomerases/metabolismo , Liases/química , Liases/genética , Liases/metabolismo , Ficobiliproteínas/metabolismo
13.
Environ Microbiol ; 24(4): 2047-2058, 2022 04.
Artigo em Inglês | MEDLINE | ID: mdl-35172392

RESUMO

Light is the crucial environmental signal for desiccation-tolerant cyanobacteria to activate photosynthesis and prepare for desiccation at dawn. However, the photobiological characteristics of desert cyanobacteria adaptation to one of the harshest habitats on Earth remain unresolved. In this study, we surveyed the genome of a subaerial desert cyanobacterium Nostoc flagelliforme and identified two phytochromes and seven cyanobacteriochromes (CBCRs) with one or more bilin-binding GAF (cGMP phosphodiesterase/adenylyl cyclase/FhlA) domains. Biochemical and spectroscopic analyses of 69 purified GAF-containing proteins from recombinant phycocyanobilin (PCB), biliverdin or phycoerythrobilin-producing Escherichia coli indicated that nine of these proteins bind chromophores. Further investigation revealed that 11 GAFs form covalent adducts responsive to near-UV and visible light: eight GAFs contained PCB chromophores, three GAFs contained biliverdin chromophores and one contained the PCB isomer, phycoviolobilin. Interestingly, COO91_03972 is the first-ever reported GAF-only CBCR capable of sensing five wavelengths of light. Bioinformatics and biochemical analyses revealed that residue P132 of COO91_03972 is essential for chromophore binding to dual-cysteine CBCRs. Furthermore, the complement of N. flagelliforme CBCRs is enriched in red light sensors. We hypothesize that these sensors are critical for the acclimatization of N. flagelliforme to weak light environments at dawn.


Assuntos
Pigmentos Biliares , Nostoc , Proteínas de Bactérias/metabolismo , Pigmentos Biliares/metabolismo , Biliverdina/metabolismo , Luz , Nostoc/genética , Nostoc/metabolismo
14.
Ann Diagn Pathol ; 58: 151911, 2022 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-35217489

RESUMO

BACKGROUND: The current WHO classification proposed high-grade biliary intraepithelial neoplasm (BilIN) and intracholecystic papillary neoplasm (ICPN) as precursors of the gallbladder carcinoma (GBC). Herein, conventional GBCs (cGBCs) were pathologically examined with respect to these two precursors. METHODS: Forty-seven cases of GBC with grossly visible invasions were collected from Fukui Saiseikai Hospital. The association of two precursors was analyzed referring to pathologic features of cGBCs and post-operative survival. RESULTS: 20 cGBCa (42.6%) were associated with either of two precursors in the surrounding mucosa: high-grade BilIN in 15 cases (31.9%) or ICPN in 5 cases (10.6%). Association of precursors was not related to gross types of and histological differentiation of cGBC. cGBCs without precursors showed frequent vascular/perineural invasion and lymph node metastasis, though cGBCs with and without precursors presented a similar post-operative survival. High-grade BilIN and ICPN associated with cGBCs showed more complicated cytoarchitectural features compared with those with no or focal invasion. CONCLUSION: More than 40% of cGBCs were associated with high-grade BilIN or ICPN, and the former was a frequent precursor. cGBCs without precursors showed aggressive pathologic features. Clinical detection of these precursors may make early treatment of cGBCs possible.


Assuntos
Adenocarcinoma , Carcinoma in Situ , Neoplasias da Vesícula Biliar , Adenocarcinoma/patologia , Pigmentos Biliares , Carcinoma in Situ/patologia , Neoplasias da Vesícula Biliar/patologia , Humanos
15.
Curr Opin Gastroenterol ; 38(2): 173-178, 2022 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-35034081

RESUMO

PURPOSE OF REVIEW: The gut microbial co-metabolism of bile-derived compounds (e.g. bile acids and bile pigments) affects colorectal cancer (CRC) risk. Here, we review recent findings with focus on selected novel aspects of bile-associated effects with interesting but unclear implications on CRC risk. RECENT FINDINGS: Numerous studies demonstrated novel biotransformation of bile acids by gut bacteria (e.g. microbial conjugation of bile acids), resulting in diverse bile acid compounds that show complex interactions with host receptors (e.g. FXR, TGR5). In addition, YAP-associated signalling in intestinal epithelial cells is modulated via bile acid receptor TGR5 and contributes to colonic tumorigenesis. Finally, studies indicate that serum levels of the bile pigment bilirubin are inversely associated with CRC risk or intestinal inflammation and that bilirubin affects gut microbiota composition. SUMMARY: Bile acids and bile pigments have multiple effects on intestinal microbe-host interactions, which may collectively modulate long-term CRC risk of the host.


Assuntos
Neoplasias Colorretais , Microbioma Gastrointestinal , Ácidos e Sais Biliares , Pigmentos Biliares , Bilirrubina , Neoplasias Colorretais/etiologia , Humanos
16.
J Phys Chem B ; 126(4): 813-821, 2022 02 03.
Artigo em Inglês | MEDLINE | ID: mdl-35076228

RESUMO

Cyanobacteriochromes (CBCRs) belong to the phytochrome superfamily of photoreceptors, the members of which utilize a linear tetrapyrrole (bilin) as a chromophore. RcaE is a representative member of a green/red-type CBCR subfamily that photoconverts between a green-absorbing dark state and red-absorbing photoproduct (Pr). Our recent crystallographic study showed that the phycocyanobilin (PCB) chromophore of RcaE adopts a unique C15-E,syn configuration in the Pr state, unlike the typical C15-E,anti configuration for the phytochromes and other CBCRs. Here, we measured Raman spectra of the Pr state of RcaE with 1064 nm excitation and explored the structure of PCB and its interacting residues under physiologically relevant aqueous conditions. We also performed measurements of RcaE in D2O as well as the sample reconstituted with the PCB labeled with 15N or with both 13C and 15N. The observed Raman spectra were analyzed by quantum mechanics/molecular mechanics (QM/MM) calculations together with molecular dynamics simulations. The Raman spectra and their isotope effects were well-reproduced by the simulated spectra of fully protonated PCB with the C15-E,syn configuration and allowed us to assign most of the observed bands. The present vibrational analysis of the all syn bilin chromophore using the QM/MM method will advance future studies on CBCRs and the related proteins by vibrational spectroscopy.


Assuntos
Fotorreceptores Microbianos , Fitocromo , Proteínas de Bactérias/química , Pigmentos Biliares/química , Simulação de Dinâmica Molecular , Fotorreceptores Microbianos/química , Fitocromo/química , Análise Espectral Raman
17.
Photosynth Res ; 151(3): 213-223, 2022 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-34564824

RESUMO

Phycobilisomes are light-harvesting antenna complexes of cyanobacteria and red algae that are comprised of chromoproteins called phycobiliproteins. PBS core structures are made up of allophycocyanin subunits. Halomicronema hongdechloris (H. hongdechloris) is one of the cyanobacteria that produce chlorophyll f (Chl f) under far-red light and is regulated by the Far-Red Light Photoacclimation gene cluster. There are five genes encoding APC in this specific gene cluster, and they are responsible for assembling the red-shifted PBS in H. hongdechloris grown under far-red light. In this study, the five apc genes located in the FaRLiP gene cluster were heterologously expressed in an Escherichia coli reconstitution system. The canonical APC-encoding genes were also constructed in the same system for comparison. Additionally, five annotated phycobiliprotein lyase-encoding genes (cpcS) from the H. hongdechloris genome were phylogenetically classified and experimentally tested for their catalytic properties including their contribution to the shifted absorption of PBS. Through analysis of recombinant proteins, we determined that the heterodimer of CpcS-I and CpcU are able to ligate a chromophore to the APC-α/APC-ß subunits. We discuss some hypotheses towards understanding the roles of the specialised APC and contributions of PBP lyases.


Assuntos
Cianobactérias , Liases , Pigmentos Biliares/metabolismo , Clorofila/análogos & derivados , Clorofila/metabolismo , Cianobactérias/metabolismo , Liases/genética , Liases/metabolismo , Ficobilissomas/metabolismo , Ficocianina/metabolismo
18.
Hepatología ; 3(2): 176-190, 2022. ilus, tab
Artigo em Espanhol | LILACS, COLNAL | ID: biblio-1396099

RESUMO

Los niveles de bilirrubina sérica normal en el adulto varían entre 0,3 mg/dL y 1,2 mg/dL, y su valor está determinado por la tasa de captación hepática, conjugación y excreción. La ictericia se hace evidente cuando los niveles de bilirrubina sérica se elevan por encima de 2,5 mg/dL a 3 mg/dL, siendo un indicador de enfermedad subyacente. La bilis es producida por los hepatocitos y fluye desde los canalículos, canales de Hering, conductos biliares intrahepáticos, conductos hepáticos derechos e izquierdos hasta llegar al duodeno. A nivel histopatológico, cualquier entidad que lleve a la acumulación intrahepática de bilis por disfunción hepatocelular u obstrucción biliar genera colestasis, que se observa en la biopsia hepática como la acumulación de tapones de color marrón verdoso de pigmento biliar en los hepatocitos, y secundariamente se observan los canalículos dilatados. Las causas de colestasis intrahepática son diversas e incluyen enfermedades como colangitis biliar primaria, colangitis esclerosante primaria, hepatitis autoinmune, hepatitis virales y toxicidad medicamentosa. Esta revisión tiene como objetivo analizar algunos tipos de hiperbilirrubinemia, resaltando sus características histopatológicas.


Normal serum bilirubin levels in adults range from 0.3 mg/dL to 1.2 mg/dL, and its value is determined by the rate of hepatic uptake, conjugation, and excretion. Jaundice becomes apparent when serum bilirubin levels rise above 2.5 mg/dL to 3.5 mg/dL and is an indicator of underlying disease. Bile is produced by hepatocytes and flows from the canaliculi, Hering's canals, intrahepatic bile ducts, and right and left hepatic ducts to the duodenum. Pathologically, any condition that leadsto intrahepatic accumulation of bile due to hepatocellular dysfunction or biliary obstruction, generates cholestasis, which is observed in liver biopsy as the accumulation of greenish-brown deposits of bile pigment in hepatocytes, with dilated canaliculi. The causes of intrahepatic cholestasis are diverse and include diseases such as primary biliary cholangitis and primary sclerosing cholangitis, autoimmune hepatitis, viral hepatitis, and drug toxicity. This review aims to analyze some types of hyperbilirubinemia, highlighting their histopathological characteristics.


Assuntos
Humanos , Patologistas , Hiperbilirrubinemia , Icterícia , Bile , Ductos Biliares Intra-Hepáticos , Pigmentos Biliares , Bilirrubina , Biópsia , Colangite Esclerosante , Colestase , Colestase Intra-Hepática , Hepatite Autoimune , Hepatite , Fígado , Cirrose Hepática Biliar
19.
Anal Methods ; 13(46): 5573-5588, 2021 12 02.
Artigo em Inglês | MEDLINE | ID: mdl-34787126

RESUMO

Faecal pigments (FPs) are ubiquitous in the environment and are a primary contaminant in groundwater and surface water. This article presents a new analytical paradigm by a fluorescence coupled extraction-based method involving FP fluorescence enhancement and minimization of background fluorescence for high sensitivity detection. FPs show higher fluorescence intensity in aliphatic alcohols due to the breaking down of higher-order H-aggregates into lower-order H-aggregates (dimers). DFT studies using the B3LYP functional and LANL2DZ basis set show π-π stacking and hydrogen-bonding contributions towards forming H-aggregated dimers of FPs in the implicit and explicit solvent environments of 1-hexanol. This study is the first report on the extractability of FPs using 1-hexanol as an efficient extraction medium in comparison to higher-order aliphatic alcohols (1-butanol, 1-hexanol and 1-octanol). Furthermore, FP-Zn(II) complexes in 1-hexanol medium significantly enhance the fluorescence emission intensity (∼14-17 times), and the emission intensity remains stable over time. This further helps to increase the detection limit of FPs in the picomolar to sub-picomolar concentration range. This study proposes a protocol involving extraction of FPs by 1-hexanol followed by the complexation of FPs with Zn(II) in the alcohol media and subsequent fluorimetric detection of the FP-Zn(II) complex with a high level of sensitivity, enabled by reduced interference from the background fluorescence of humic acid. The complexation behaviour of FPs with various metal salts was also examined, which provided an understanding of the fluorescence behaviour of FPs with various other metal ions commonly present in natural environmental water. The proposed analytical method has been further validated using real water samples.


Assuntos
Etanol , Urobilina , Pigmentos Biliares , Fluorometria , Solventes
20.
Chem Rev ; 121(24): 14906-14956, 2021 12 22.
Artigo em Inglês | MEDLINE | ID: mdl-34669383

RESUMO

This review adds the bilin-binding phytochromes to the Chemical Reviews thematic issue "Optogenetics and Photopharmacology". The work is structured into two parts. We first outline the photochemistry of the covalently bound tetrapyrrole chromophore and summarize relevant spectroscopic, kinetic, biochemical, and physiological properties of the different families of phytochromes. Based on this knowledge, we then describe the engineering of phytochromes to further improve these chromoproteins as photoswitches and review their employment in an ever-growing number of different optogenetic applications. Most applications rely on the light-controlled complex formation between the plant photoreceptor PhyB and phytochrome-interacting factors (PIFs) or C-terminal light-regulated domains with enzymatic functions present in many bacterial and algal phytochromes. Phytochrome-based optogenetic tools are currently implemented in bacteria, yeast, plants, and animals to achieve light control of a wide range of biological activities. These cover the regulation of gene expression, protein transport into cell organelles, and the recruitment of phytochrome- or PIF-tagged proteins to membranes and other cellular compartments. This compilation illustrates the intrinsic advantages of phytochromes compared to other photoreceptor classes, e.g., their bidirectional dual-wavelength control enabling instant ON and OFF regulation. In particular, the long wavelength range of absorption and fluorescence within the "transparent window" makes phytochromes attractive for complex applications requiring deep tissue penetration or dual-wavelength control in combination with blue and UV light-sensing photoreceptors. In addition to the wide variability of applications employing natural and engineered phytochromes, we also discuss recent progress in the development of bilin-based fluorescent proteins.


Assuntos
Pigmentos Biliares , Fitocromo , Animais , Pigmentos Biliares/química , Luz , Optogenética , Fotoquímica , Células Fotorreceptoras/metabolismo , Fitocromo/química
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA
...