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1.
BMJ Open Qual ; 12(3)2023 07.
Artigo em Inglês | MEDLINE | ID: mdl-37463784

RESUMO

OBJECTIVE: Missed or cancelled imaging tests may be invisible to the ordering clinician and result in diagnostic delay. We developed an outpatient results notification tool (ORNT) to alert physicians of patients' missed radiology studies. DESIGN: Randomised controlled evaluation of a quality improvement intervention. SETTING: 23 primary care and subspecialty ambulatory clinics at an urban academic medical centre. PARTICIPANTS: 276 physicians randomised to intervention or usual care. MAIN OUTCOME MEASURE: 90-day test completion of missed imaging tests. RESULTS: We included 3675 radiology tests in our analysis: 1769 ordered in the intervention group and 1906 in the usual care group. A higher per cent of studies were completed for intervention compared with usual care groups in CT (20.7% vs 15.3%, p=0.06), general radiology (19.6% vs 12.0%, p=0.02) and, in aggregate, across all modalities (18.1% vs 16.1%, p=0.03). In the multivariable regression model adjusting for sex, age and insurance type and accounting for clustering with random effects at the level of the physician, the intervention group had a 36% greater odds of test completion than the usual care group (OR: 1.36 (1.097-1.682), p=0.005). In the Cox regression model, patients in the intervention group were 1.32 times more likely to complete their test in a timely fashion (HR: 1.32 (1.10-1.58), p=0.003). CONCLUSIONS: An electronic alert that notified the responsible clinician of a missed imaging test ordered in an ambulatory clinic reduced the number of incomplete tests at 90 days. Further study of the obstacles to completing recommended diagnostic testing may allow for the development of better tools to support busy clinicians and their patients and reduce the risk of diagnostic delays.


Assuntos
Diagnóstico Tardio , Diagnóstico por Imagem , Perda de Seguimento , Humanos , Assistência Ambulatorial , Médicos
2.
Oncol Lett ; 25(4): 173, 2023 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-36970608

RESUMO

Prostate cancer (PCa) is one of the most prevalent types of cancer in men worldwide; however, the main diagnostic tests available for PCa have limitations and a biopsy is required for histopathological confirmation of the disease. Prostate-specific antigen (PSA) is the main biomarker used for the early detection of PCa, but an elevated serum concentration is not cancer-specific. Therefore, there is a need for the discovery of new non-invasive biomarkers that can accurately diagnose PCa. The present study used trichloroacetic acid-induced protein precipitation and liquid chromatography-mass spectrometry to profile endogenous peptides in urine samples from patients with PCa (n=33), benign prostatic hyperplasia (n=25) and healthy individuals (n=28). Receiver operating characteristic curve analysis was performed to evaluate the diagnostic performance of urinary peptides. In addition, Proteasix tool was used for in silico prediction of protease cleavage sites. Five urinary peptides derived from uromodulin were revealed to be significantly altered between the study groups, all of which were less abundant in the PCa group. This peptide panel showed a high potential to discriminate between the study groups, resulting in area under the curve (AUC) values between 0.788 and 0.951. In addition, urinary peptides outperformed PSA in discriminating between malignant and benign prostate conditions (AUC=0.847), showing high sensitivity (81.82%) and specificity (88%). From in silico analyses, the proteases HTRA2, KLK3, KLK4, KLK14 and MMP25 were identified as potentially involved in the degradation of uromodulin peptides in the urine of patients with PCa. In conclusion, the present study allowed the identification of urinary peptides with potential for use as non-invasive biomarkers in PCa diagnosis.

3.
J Am Chem Soc ; 144(14): 6237-6250, 2022 04 13.
Artigo em Inglês | MEDLINE | ID: mdl-35362954

RESUMO

Chronically elevated circulating fatty acid levels promote lipid accumulation in nonadipose tissues and cause lipotoxicity. Adipose triglyceride lipase (ATGL) critically determines the release of fatty acids from white adipose tissue, and accumulating evidence suggests that inactivation of ATGL has beneficial effects on lipotoxicity-driven disorders including insulin resistance, steatohepatitis, and heart disease, classifying ATGL as a promising drug target. Here, we report on the development and biological characterization of the first small-molecule inhibitor of human ATGL. This inhibitor, designated NG-497, selectively inactivates human and nonhuman primate ATGL but not structurally and functionally related lipid hydrolases. We demonstrate that NG-497 abolishes lipolysis in human adipocytes in a dose-dependent and reversible manner. The combined analysis of mouse- and human-selective inhibitors, chimeric ATGL proteins, and homology models revealed detailed insights into enzyme-inhibitor interactions. NG-497 binds ATGL within a hydrophobic cavity near the active site. Therein, three amino acid residues determine inhibitor efficacy and species selectivity and thus provide the molecular scaffold for selective inhibition.


Assuntos
Aciltransferases/antagonistas & inibidores , Adipócitos , Ácidos Graxos/metabolismo , Lipólise , Aciltransferases/metabolismo , Adipócitos/metabolismo , Animais , Humanos , Lipólise/fisiologia , Camundongos
4.
Food Chem ; 381: 132170, 2022 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-35121327

RESUMO

The market for plant protein-based substitutes for cheeses is growing, but the sensory properties are distinctively different from the original products. Hence, natural and vegan cheesy flavors are needed to aromatize the products. A cheesy, sweaty and parmesan-like aroma was produced by fermentation of soy drink with Agrocybe aegerita. Aroma dilution analysis revealed short-chain fatty acids (SCFAs) as main influencing cheesy odorants analyzed by gas chromatography-mass spectrometry-olfactometry. In comparison to the five cheese varieties, the SCFA profile of the fermented soy drink revealed similarities with Parmesan and Emmental cheese. Meanwhile, principal component analysis showed an approximation of the aroma profile after fermentation with A. aegerita to those of cheeses. 3-Methylbutanoic acid was synthesized from the protein fraction, while the oil fraction contributed to the formation of unbranched SCFAs like butanoic acid. Accordingly, the production of these compounds can be increased by addition of the fractions.


Assuntos
Queijo , Compostos Orgânicos Voláteis , Agrocybe , Queijo/análise , Fermentação , Odorantes , Paladar , Compostos Orgânicos Voláteis/análise
5.
Food Chem ; 378: 131956, 2022 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-35045369

RESUMO

When we are trying to decrease caloric intake by reducing fat content, aroma perception of the food is changed as well. Since previous studies indicated that foaming changes the aroma release, it was our goal to understand the physico-chemical background of this effect. Therefore, ten aroma compounds were added to a foamed acidified dairy matrix (4% milk protein, 1% gelatin, 60% gas volume). We simulated oral temperature conditions in a simplistic way through incubation at 40 °C and analyzed aroma release using headspace-solid phase microextraction-gas chromatography-ion mobility spectrometry. Significantly more highly hydrophobic aroma compounds were released from the foamed matrix than the unfoamed matrix, while compounds of intermediate hydrophobicity were released more from unfoamed matrix. The effect was independent from foam collapse and persisted for hours afterwards. Analytical results were complemented by orthonasal and retronasal sensory perception studies, which confirmed significant differences between aroma release behavior from foamed foods.


Assuntos
Odorantes , Compostos Orgânicos Voláteis , Cromatografia Gasosa-Espectrometria de Massas , Odorantes/análise , Percepção , Microextração em Fase Sólida , Compostos Orgânicos Voláteis/análise
6.
Talanta ; 229: 122251, 2021 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-33838768

RESUMO

A detailed molecular flavor profile was necessary to understand the low acceptance of soy drink by Western consumers. Accordingly, key odor-active compounds were detected by means of aroma dilution analyses coupled with gas chromatography-mass spectrometry-olfactometry after application of suitable solvent-free volatile extraction techniques. Four quantification methods (standard addition, external calibration, internal standard, and stable isotope dilution assay) were developed and validated to measure the concentrations after direct immersion-stir bar sorptive extraction. The quantitative methods provided correctness between 97% and 111% and precision ranging from 78% to 99% for the 21 key odorants. Considering the advantages to be efficient, easy to perform and cheap, internal standard method was further applied to four commercially available soy drinks in Germany. Correlated to a sensory acceptance test (n = 52) contents of 1-octen-3-one, (E,E)-2,4-decadienal and 2-methoxy-4-vinylphenol were suggested to be linked to the aversion of Western consumers to soy drink.

7.
Bioorg Med Chem ; 28(16): 115610, 2020 08 15.
Artigo em Inglês | MEDLINE | ID: mdl-32690265

RESUMO

High serum fatty acid (FA) levels are causally linked to the development of insulin resistance, which eventually progresses to type 2 diabetes and non-alcoholic fatty liver disease (NAFLD) generalized in the term metabolic syndrome. Adipose triglyceride lipase (ATGL) is the initial enzyme in the hydrolysis of intracellular triacylglycerol (TG) stores, liberating fatty acids that are released from adipocytes into the circulation. Hence, ATGL-specific inhibitors have the potential to lower circulating FA concentrations, and counteract the development of insulin resistance and NAFLD. In this article, we report about structure-activity relationship (SAR) studies of small molecule inhibitors of murine ATGL which led to the development of Atglistatin. Atglistatin is a specific inhibitor of murine ATGL, which has proven useful for the validation of ATGL as a potential drug target.


Assuntos
Inibidores Enzimáticos/química , Inibidores Enzimáticos/farmacologia , Lipase/antagonistas & inibidores , Compostos de Fenilureia/química , Compostos de Fenilureia/farmacologia , Animais , Descoberta de Drogas , Lipase/química , Lipase/metabolismo , Lipólise/efeitos dos fármacos , Camundongos , Relação Estrutura-Atividade , Triglicerídeos/sangue
9.
Nat Commun ; 8: 14859, 2017 03 22.
Artigo em Inglês | MEDLINE | ID: mdl-28327588

RESUMO

Elevated circulating fatty acids (FAs) contribute to the development of obesity-associated metabolic complications such as insulin resistance (IR) and non-alcoholic fatty liver disease (NAFLD). Hence, reducing adipose tissue lipolysis to diminish the mobilization of FAs and lower their respective plasma concentrations represents a potential treatment strategy to counteract obesity-associated disorders. Here we show that specific inhibition of adipose triglyceride lipase (Atgl) with the chemical inhibitor, Atglistatin, effectively reduces adipose tissue lipolysis, weight gain, IR and NAFLD in mice fed a high-fat diet. Importantly, even long-term treatment does not lead to lipid accumulation in ectopic tissues such as the skeletal muscle or heart. Thus, the severe cardiac steatosis and cardiomyopathy that is observed in genetic models of Atgl deficiency does not occur in Atglistatin-treated mice. Our data validate the pharmacological inhibition of Atgl as a potentially powerful therapeutic strategy to treat obesity and associated metabolic disorders.


Assuntos
Tecido Adiposo Branco/enzimologia , Fígado Gorduroso/tratamento farmacológico , Fígado Gorduroso/enzimologia , Resistência à Insulina , Lipase/antagonistas & inibidores , Animais , Peso Corporal/efeitos dos fármacos , Dieta Hiperlipídica , Fígado Gorduroso/sangue , Fígado Gorduroso/prevenção & controle , Comportamento Alimentar , Glucose/metabolismo , Homeostase/efeitos dos fármacos , Humanos , Lipase/metabolismo , Lipólise/efeitos dos fármacos , Masculino , Camundongos Endogâmicos C57BL , Obesidade/tratamento farmacológico , Obesidade/enzimologia , Obesidade/patologia , Compostos de Fenilureia/farmacologia , Compostos de Fenilureia/uso terapêutico
10.
Biochim Biophys Acta ; 1861(5): 462-70, 2016 May.
Artigo em Inglês | MEDLINE | ID: mdl-26869448

RESUMO

Monoglyceride lipases (MGLs) are a group of α/ß-hydrolases that catalyze the hydrolysis of monoglycerides (MGs) into free fatty acids and glycerol. This reaction serves different physiological functions, namely in the last step of phospholipid and triglyceride degradation, in mammalian endocannabinoid and arachidonic acid metabolism, and in detoxification processes in microbes. Previous crystal structures of MGLs from humans and bacteria revealed conformational plasticity in the cap region of this protein and gave insight into substrate binding. In this study, we present the structure of a MGL from Saccharomyces cerevisiae called Yju3p in its free form and in complex with a covalently bound substrate analog mimicking the tetrahedral intermediate of MG hydrolysis. These structures reveal a high conservation of the overall shape of the MGL cap region and also provide evidence for conformational changes in the cap of Yju3p. The complex structure reveals that, despite the high structural similarity, Yju3p seems to have an additional opening to the substrate binding pocket at a different position compared to human and bacterial MGL. Substrate specificities towards MGs with saturated and unsaturated alkyl chains of different lengths were tested and revealed highest activity towards MG containing a C18:1 fatty acid.


Assuntos
Monoacilglicerol Lipases/química , Monoglicerídeos/química , Proteínas de Saccharomyces cerevisiae/química , Sítios de Ligação , Catálise , Clonagem Molecular , Cristalização , Hidrólise , Simulação de Dinâmica Molecular , Monoacilglicerol Lipases/genética , Monoacilglicerol Lipases/metabolismo , Monoglicerídeos/metabolismo , Mutagênese Sítio-Dirigida , Mutação , Ligação Proteica , Conformação Proteica , Proteínas Recombinantes/química , Proteínas Recombinantes/metabolismo , Proteínas de Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/metabolismo , Relação Estrutura-Atividade , Especificidade por Substrato
11.
J Biol Chem ; 290(30): 18438-53, 2015 Jul 24.
Artigo em Inglês | MEDLINE | ID: mdl-25953897

RESUMO

The coordinated breakdown of intracellular triglyceride (TG) stores requires the exquisitely regulated interaction of lipolytic enzymes with regulatory, accessory, and scaffolding proteins. Together they form a dynamic multiprotein network designated as the "lipolysome." Adipose triglyceride lipase (Atgl) catalyzes the initiating step of TG hydrolysis and requires comparative gene identification-58 (Cgi-58) as a potent activator of enzyme activity. Here, we identify adipocyte-type fatty acid-binding protein (A-Fabp) and other members of the fatty acid-binding protein (Fabp) family as interaction partners of Cgi-58. Co-immunoprecipitation, microscale thermophoresis, and solid phase assays proved direct protein/protein interaction between A-Fabp and Cgi-58. Using nuclear magnetic resonance titration experiments and site-directed mutagenesis, we located a potential contact region on A-Fabp. In functional terms, A-Fabp stimulates Atgl-catalyzed TG hydrolysis in a Cgi-58-dependent manner. Additionally, transcriptional transactivation assays with a luciferase reporter system revealed that Fabps enhance the ability of Atgl/Cgi-58-mediated lipolysis to induce the activity of peroxisome proliferator-activated receptors. Our studies identify Fabps as crucial structural and functional components of the lipolysome.


Assuntos
1-Acilglicerol-3-Fosfato O-Aciltransferase/metabolismo , Proteínas de Ligação a Ácido Graxo/metabolismo , Lipase/metabolismo , Complexos Multiproteicos/metabolismo , Triglicerídeos/metabolismo , 1-Acilglicerol-3-Fosfato O-Aciltransferase/genética , Tecido Adiposo/metabolismo , Animais , Células COS , Chlorocebus aethiops , Proteínas de Ligação a Ácido Graxo/genética , Humanos , Ligantes , Lipase/genética , Lipólise/genética , Lipossomos/metabolismo , Camundongos , Complexos Multiproteicos/genética , Receptores Ativados por Proliferador de Peroxissomo/metabolismo , Proteólise
12.
Bioorg Med Chem ; 23(12): 2904-16, 2015 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-25778769

RESUMO

Adipose triglyceride lipase (ATGL) catalyzes the degradation of cellular triacylglycerol stores and strongly determines the concentration of circulating fatty acids (FAs). High serum FA levels are causally linked to the development of insulin resistance and impaired glucose tolerance, which eventually progresses to overt type 2 diabetes. ATGL-specific inhibitors could be used to lower circulating FAs, which can counteract the development of insulin resistance. In this article, we report about structure-activity relationship (SAR) studies of small molecule inhibitors of ATGL based on a hydrazone chemotype. The SAR indicated that the binding pocket of ATGL requests rather linear compounds without bulky substituents. The best inhibitor showed an IC50=10µM in an assay with COS7-cell lysate overexpressing murine ATGL.


Assuntos
Inibidores Enzimáticos/química , Inibidores Enzimáticos/farmacologia , Hidrazonas/química , Hidrazonas/farmacologia , Lipase/antagonistas & inibidores , Animais , Células COS , Chlorocebus aethiops , Inibidores Enzimáticos/síntese química , Hidrazonas/síntese química , Lipase/metabolismo , Metabolismo dos Lipídeos/efeitos dos fármacos , Camundongos , Bibliotecas de Moléculas Pequenas/síntese química , Bibliotecas de Moléculas Pequenas/química , Bibliotecas de Moléculas Pequenas/farmacologia , Relação Estrutura-Atividade , Triglicerídeos/metabolismo
13.
Nat Chem Biol ; 9(12): 785-7, 2013 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-24096302

RESUMO

Adipose triglyceride lipase (ATGL) is rate limiting in the mobilization of fatty acids from cellular triglyceride stores. This central role in lipolysis marks ATGL as an interesting pharmacological target as deregulated fatty acid metabolism is closely linked to dyslipidemic and metabolic disorders. Here we report on the development and characterization of a small-molecule inhibitor of ATGL. Atglistatin is selective for ATGL and reduces fatty acid mobilization in vitro and in vivo.


Assuntos
Lipase/antagonistas & inibidores , Lipase/metabolismo , Compostos de Fenilureia/farmacologia , Tecido Adiposo Branco , Animais , Regulação Enzimológica da Expressão Gênica , Concentração Inibidora 50 , Lipase/genética , Camundongos , Camundongos Knockout , Estrutura Molecular
14.
J Biol Chem ; 288(43): 31093-104, 2013 Oct 25.
Artigo em Inglês | MEDLINE | ID: mdl-24014019

RESUMO

Monoacylglycerol lipases (MGLs) play an important role in lipid catabolism across all kingdoms of life by catalyzing the release of free fatty acids from monoacylglycerols. The three-dimensional structures of human and a bacterial MGL were determined only recently as the first members of this lipase family. In addition to the α/ß-hydrolase core, they showed unexpected structural similarities even in the cap region. Nevertheless, the structural basis for substrate binding and conformational changes of MGLs is poorly understood. Here, we present a comprehensive study of five crystal structures of MGL from Bacillus sp. H257 in its free form and in complex with different substrate analogs and the natural substrate 1-lauroylglycerol. The occurrence of different conformations reveals a high degree of conformational plasticity of the cap region. We identify a specific residue, Ile-145, that might act as a gatekeeper restricting access to the binding site. Site-directed mutagenesis of Ile-145 leads to significantly reduced hydrolase activity. Bacterial MGLs in complex with 1-lauroylglycerol, myristoyl, palmitoyl, and stearoyl substrate analogs enable identification of the binding sites for the alkyl chain and the glycerol moiety of the natural ligand. They also provide snapshots of the hydrolytic reaction of a bacterial MGL at different stages. The alkyl chains are buried in a hydrophobic tunnel in an extended conformation. Binding of the glycerol moiety is mediated via Glu-156 and water molecules. Analysis of the structural features responsible for cap plasticity and the binding modes of the ligands suggests conservation of these features also in human MGL.


Assuntos
Bacillus/enzimologia , Proteínas de Bactérias/química , Monoacilglicerol Lipases/química , Monoglicerídeos/química , Bacillus/genética , Proteínas de Bactérias/genética , Cristalografia por Raios X , Humanos , Hidrólise , Monoacilglicerol Lipases/genética , Monoglicerídeos/genética , Mutagênese Sítio-Dirigida , Estrutura Terciária de Proteína , Relação Estrutura-Atividade , Especificidade por Substrato
15.
Cereb Cortex ; 21(8): 1771-82, 2011 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-21139075

RESUMO

Cortical γ-aminobutyric acid (GABA)ergic interneurons in rodents originate mainly in ventrally positioned ganglionic eminences (GEs), but their origin in primates is still debated. We studied human fetal forebrains during the first half of gestation (5-23 gestational weeks, gw) for the expression of ventral transcription factors, Nkx2.1, Dlx1,2, Lhx6, and Mash1, important for development of neocortical interneurons. In embryonic (5-8 gw) human forebrain, these factors were expressed in the GE but also dorsally in the neocortical ventricular/subventricular zones (VZ/SVZ). Furthermore, their expression was retained in cells of all fetal cortical layers up to midgestation (20 gw). Nkx2.1 continued to be expressed not only in the GE but also in a subpopulation of neocortical interneurons. Moreover, proliferation marker Ki67 revealed that calretinin(+), Mash1(+), and Nkx2.1(+) cells proliferate in the neocortical VZ/SVZ at midgestation. At least some of the Mash1(+) progenitors in the neocortical SVZ could be colabeled with GABA, whereas others were oligodendrocyte progenitors, indicating a link between the 2 lineages. Taken together, these results suggest the existence of several categories of dorsal interneuronal progenitors in the human neocortical VZ/SVZ, in addition to ventrally derived cortical interneurons described in rodents. These human-specific developmental events may underlie human brain's higher complexity and capacity to process information.


Assuntos
Interneurônios/metabolismo , Neocórtex/embriologia , Neocórtex/metabolismo , Neurogênese/fisiologia , Fatores de Transcrição/biossíntese , Diferenciação Celular/fisiologia , Linhagem da Célula/fisiologia , Feminino , Feto/citologia , Feto/fisiologia , Idade Gestacional , Humanos , Interneurônios/citologia , Masculino , Neocórtex/citologia , Células-Tronco Neurais/citologia , Células-Tronco Neurais/metabolismo , Fatores de Transcrição/genética , Ácido gama-Aminobutírico/fisiologia
16.
Pers Soc Psychol Bull ; 36(4): 443-54, 2010 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-20363901

RESUMO

Three studies explored think ("I think . . . ") versus feel ("I feel . . . ") message framing effects on persuasion.The authors propose a matching hypothesis, suggesting that think framing will be more persuasive when the target attitude or message recipient is cognitively oriented, whereas feel framing will be more persuasive when the target attitude or message recipient is affectively oriented. Study 1 presented cognitively and affectively oriented individuals with a think- or feel-framed message. Study 2 primed cognitive or affective orientation and then presented a think- or feel-framed message. Study 3 presented male and female participants with an advertisement containing think- or feel-framed arguments. Results indicated that think (feel) framing was more persuasive when the target attitude or recipient was cognitively (affectively) oriented. Moreover, Study 2 demonstrated that this matching effect was mediated by processing fluency. Theoretical and practical implications are discussed.


Assuntos
Emoções , Comunicação Persuasiva , Pensamento , Feminino , Humanos , Masculino
17.
J Biol Chem ; 285(15): 11078-86, 2010 Apr 09.
Artigo em Inglês | MEDLINE | ID: mdl-20106966

RESUMO

The concentration of the second messenger cAMP is tightly controlled in cells by the activity of phosphodiesterases. We have previously described how the protein kinase A-anchoring protein mAKAP serves as a scaffold for the cAMP-dependent protein kinase PKA and the cAMP-specific phosphodiesterase PDE4D3 in cardiac myocytes. PKA and PDE4D3 constitute a negative feedback loop whereby PKA-catalyzed phosphorylation and activation of PDE4D3 attenuate local cAMP levels. We now show that protein phosphatase 2A (PP2A) associated with mAKAP complexes is responsible for reversing the activation of PDE4D3 by catalyzing the dephosphorylation of PDE4D3 serine residue 54. Mapping studies reveal that a C-terminal mAKAP domain (residues 2085-2319) binds PP2A. Binding to mAKAP is required for PP2A function, such that deletion of the C-terminal domain enhances both base-line and forskolin-stimulated PDE4D3 activity. Interestingly, PP2A holoenzyme associated with mAKAP complexes in the heart contains the PP2A targeting subunit B56delta. Like PDE4D3, B56delta is a PKA substrate, and PKA phosphorylation of mAKAP-bound B56delta enhances phosphatase activity 2-fold in the complex. Accordingly, expression of a B56delta mutant that cannot be phosphorylated by PKA results in increased PDE4D3 phosphorylation. Taken together, our findings demonstrate that PP2A associated with mAKAP complexes promotes PDE4D3 dephosphorylation, serving both to inhibit PDE4D3 in unstimulated cells and also to mediate a cAMP-induced positive feedback loop following adenylyl cyclase activation and B56delta phosphorylation. In general, PKA.PP2A.mAKAP complexes exemplify how protein kinases and phosphatases may participate in molecular signaling complexes to dynamically regulate localized intracellular signaling.


Assuntos
Proteínas de Ancoragem à Quinase A/metabolismo , Nucleotídeo Cíclico Fosfodiesterase do Tipo 4/metabolismo , Regulação Enzimológica da Expressão Gênica , Proteína Fosfatase 2/química , Animais , Linhagem Celular , AMP Cíclico/metabolismo , Retroalimentação Fisiológica , Humanos , Fosforilação , Ligação Proteica , Mapeamento de Interação de Proteínas , Proteína Fosfatase 2/metabolismo , Estrutura Terciária de Proteína , Ratos , Transdução de Sinais
18.
Biochem J ; 381(Pt 3): 587-92, 2004 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-15182229

RESUMO

mAKAP (muscle-selective A-kinase-anchoring protein) co-ordinates a cAMP-sensitive negative-feedback loop comprising PKA (cAMP-dependent protein kinase) and the cAMP-selective PDE4D3 (phosphodiesterase 4D3). In vitro and cellular experiments demonstrate that PKA-phosphorylation of PDE4D3 on Ser-13 increases the affinity of PDE4D3 for mAKAP. Our data suggest that activation of mAKAP-anchored PKA enhances the recruitment of PDE4D3, allowing for quicker signal termination.


Assuntos
3',5'-AMP Cíclico Fosfodiesterases/metabolismo , Proteínas Adaptadoras de Transdução de Sinal/fisiologia , Proteínas Quinases Dependentes de AMP Cíclico/metabolismo , Sítios de Ligação , Nucleotídeo Cíclico Fosfodiesterase do Tipo 4 , Ativação Enzimática/fisiologia , Humanos , Rim/citologia , Rim/embriologia , Rim/enzimologia , Mimetismo Molecular/fisiologia , Fragmentos de Peptídeos/metabolismo , Peptídeos/metabolismo , Fosforilação , Ligação Proteica/fisiologia , Mapeamento de Interação de Proteínas , Serina/metabolismo
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