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1.
Parkinsonism Relat Disord ; 91: 128-134, 2021 10.
Artigo em Inglês | MEDLINE | ID: mdl-34607089

RESUMO

BACKGROUND: Impaired bioenergetics are partially involved in the pathogenesis of Parkinson's disease (PD). Phosphoglycerate kinase (PGK), an essential enzyme for glycolysis, has recently attracted attention due to its pathogenic role in PD and as a target for disease-modifying therapies. This study is aimed to evaluate the profiles of PGK activity in red blood cells (RBCs) of PD patients and controls. METHODS: Sixty-eight PD patients and thirty-four age-matched unrelated controls were enrolled. PGK activities of RBCs were measured by the established colorimetric assay and standardized by the same RBC samples. RESULTS: PGK activity of the PD group was significantly higher than that of the control group in participants aged sixty-five years or younger, whereas it was not significantly different between the two groups at any age. PGK activity was positively correlated with aging in the control group, but this was not noted in the PD group. On multivariable analysis by partial correlation in the PD group, PGK activity was negatively correlated with the specific binding ratio of dopamine transporter scintigraphy in the striatum. The levodopa-equivalent daily dose was not significantly correlated with the enzyme activity. CONCLUSION: The results support the following: 1) elevation of PGK activities in RBCs can be detected in relatively young PD patients and with normal aging; 2) the degree of striatonigral degeneration is associated with elevated PGK activities. These are important considerations when the PGK assay is applied as a diagnostic biomarker of PD and to therapeutically monitor PGK-enhancing treatments.


Assuntos
Envelhecimento/sangue , Eritrócitos/enzimologia , Doença de Parkinson/enzimologia , Fosfoglicerato Quinase/sangue , Idoso , Idoso de 80 Anos ou mais , Estudos de Casos e Controles , Feminino , Glicólise , Humanos , Masculino , Pessoa de Meia-Idade
2.
Neurology ; 91(11): e1077-e1082, 2018 09 11.
Artigo em Inglês | MEDLINE | ID: mdl-30111548

RESUMO

OBJECTIVE: To study the variable clinical picture and exercise tolerance of patients with phosphoglycerate kinase (PGK) 1 deficiency and how it relates to residual PGK enzyme activity. METHODS: In this case series study, we evaluated 7 boys and men from 5 families with PGK1 deficiency. Five had pure muscle symptoms, while 2 also had mild intellectual disability with or without anemia. Muscle glycolytic and oxidative capacities were evaluated by an ischemic forearm exercise test and by cycle ergometry. RESULTS: Enzyme levels of PGK were 4% to 9% of normal in red cells and 5% to10% in muscle in pure myopathy patients and 2.6% in both muscle and red cells in the 2 patients with multisystem involvement. Patients with pure myopathy had greater increases in lactate with ischemic exercise (2-3 mmol/L) vs the 2 multisystem-affected patients (<1 mmol/L). Myopathy patients had higher oxidative capacity in cycle exercise vs multisystem affected patients (≈30 vs ≈15 mL/kg per minute). One multisystem-affected patient developed frank myoglobinuria after the short exercise test. CONCLUSIONS: This case series study of PGK1 deficiency suggests that the level of impaired glycolysis in PGK deficiency is a major determinant of phenotype. Lower glycolytic capacity in PGK1 deficiency seems to result in multisystem involvement and increased susceptibility to exertional rhabdomyolysis.


Assuntos
Tolerância ao Exercício/fisiologia , Doenças Genéticas Ligadas ao Cromossomo X/enzimologia , Doenças Genéticas Ligadas ao Cromossomo X/fisiopatologia , Erros Inatos do Metabolismo/enzimologia , Erros Inatos do Metabolismo/fisiopatologia , Fosfoglicerato Quinase/deficiência , Fosfoglicerato Quinase/metabolismo , Ergometria , Teste de Esforço , Doenças Genéticas Ligadas ao Cromossomo X/complicações , Doenças Genéticas Ligadas ao Cromossomo X/diagnóstico , Humanos , Deficiência Intelectual/sangue , Deficiência Intelectual/complicações , Deficiência Intelectual/enzimologia , Deficiência Intelectual/fisiopatologia , Ácido Láctico/sangue , Masculino , Erros Inatos do Metabolismo/complicações , Erros Inatos do Metabolismo/diagnóstico , Músculo Esquelético/metabolismo , Doenças Musculares/sangue , Doenças Musculares/complicações , Doenças Musculares/enzimologia , Doenças Musculares/fisiopatologia , Fenótipo , Fosfoglicerato Quinase/sangue
3.
Inflamm Res ; 65(10): 815-25, 2016 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-27342824

RESUMO

BACKGROUND: Some studies have indicated that glucose metabolism plays an important role in the pathogenesis of rheumatoid arthritis (RA). This study aimed to find the novel genes affecting glucose metabolism in RA. MATERIALS/METHODS: Synovial tissues of collagen-induced arthritis (CIA) were analyzed with Rat Glucose Metabolism RT(2) Profiler™ PCR Array to screen those genes with special expressions in glucose metabolism. Real-time PCR, western blotting, and ELISA were used to confirm the result in synovial tissues and blood of human RA. Culture synovial fibroblast cells (RASF) was treated with siRNA to suppress expressions of the target genes. CCK-8 cell proliferation assay and two-compartment transwell system were performed to examine cell proliferation and cell migration of the treated RASF. RESULTS: Both PCR array and real-time PCR detected the up-regulation of ENO1, HK2, and PGK1 and the down-regulation of PCK1 and PDK4 in synovial tissues of CIA rats. Real-time PCR and western blotting detected the increased expression of ENO1 and PGK1 in RA synovial tissues. ELISA detected a high level of PGK1 in the blood of RA patients. Decreased cell proliferation and cell migration capabilities were significantly detected in RASF following treatment of anti-PGK1 siRNA. IL-1ß and IFN-γ rather than TNF-α and IL-1α levels were significantly declined in supernatants of the treated RASF. CONCLUSIONS: PGK1, a glycolytic enzyme catalyzing the conversion of 3-phosphoglycerate into 2-phosphoglycerate, has increased expression in synovial tissues and blood of RA, which may be involved in pro-inflammation and synovial hyperplasia of the disease.


Assuntos
Artrite Reumatoide/metabolismo , Fosfoglicerato Quinase/metabolismo , Membrana Sinovial/metabolismo , Adulto , Idoso , Animais , Artrite , Artrite Experimental/genética , Artrite Experimental/metabolismo , Artrite Reumatoide/sangue , Artrite Reumatoide/genética , Biomarcadores Tumorais/sangue , Biomarcadores Tumorais/genética , Biomarcadores Tumorais/metabolismo , Colágeno , Citocinas/metabolismo , Proteínas de Ligação a DNA/sangue , Proteínas de Ligação a DNA/genética , Proteínas de Ligação a DNA/metabolismo , Feminino , Glucose/metabolismo , Humanos , Peptídeos e Proteínas de Sinalização Intracelular/genética , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Cinesinas/sangue , Cinesinas/genética , Cinesinas/metabolismo , Masculino , Pessoa de Meia-Idade , Fosfoenolpiruvato Carboxiquinase (GTP)/genética , Fosfoenolpiruvato Carboxiquinase (GTP)/metabolismo , Fosfoglicerato Quinase/sangue , Fosfoglicerato Quinase/genética , Fosfopiruvato Hidratase/sangue , Fosfopiruvato Hidratase/genética , Fosfopiruvato Hidratase/metabolismo , Proteínas Serina-Treonina Quinases/sangue , Proteínas Serina-Treonina Quinases/genética , Proteínas Serina-Treonina Quinases/metabolismo , Piruvato Desidrogenase Quinase de Transferência de Acetil , Ratos Wistar , Proteínas Supressoras de Tumor/sangue , Proteínas Supressoras de Tumor/genética , Proteínas Supressoras de Tumor/metabolismo , Adulto Jovem
4.
Hepatology ; 56(6): 2231-41, 2012 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-22706893

RESUMO

UNLABELLED: Survival of patients with hepatocellular carcinoma (HCC) remains poor, which is largely attributed to active angiogenesis. However, the mechanisms underlying angiogenesis in HCC remain to be discovered. In this study, we found that long noncoding RNA associated with microvascular invasion in HCC (lncRNA MVIH) (lncRNA associated with microvascular invasion in HCC) was generally overexpressed in HCC. In a cohort of 215 HCC patients, the overexpression of MVIH was associated with frequent microvascular invasion (P = 0.016) and a higher tumor node metastasis stage (P = 0.009) as well as decreased recurrence-free survival (RFS) (P < 0.001) and overall survival (P = 0.007). Moreover, the up-regulation of MVIH served as an independent risk factor to predict poor RFS. We also found that MVIH could promote tumor growth and intrahepatic metastasis by activating angiogenesis in mouse models. Subsequent investigations indicated that MVIH could activate tumor-inducing angiogenesis by inhibiting the secretion of phosphoglycerate kinase 1 (PGK1). Additionally, in 65 HCC samples, MVIH expression was inversely correlated with the serum level of PGK1 and positively correlated with the microvessel density. CONCLUSION: Deregulation of lncRNA MVIH is a predictor for poor RFS of HCC patients after hepatectomy and could be utilized as a potential target for new adjuvant therapies against active angiogenesis.


Assuntos
Carcinoma Hepatocelular/irrigação sanguínea , Carcinoma Hepatocelular/genética , Neoplasias Hepáticas/irrigação sanguínea , Neoplasias Hepáticas/genética , Neovascularização Patológica/genética , Fosfoglicerato Quinase/metabolismo , RNA Longo não Codificante/genética , Animais , Carcinoma Hepatocelular/patologia , Linhagem Celular Tumoral , Intervalo Livre de Doença , Feminino , Expressão Gênica , Hepatectomia , Humanos , Estimativa de Kaplan-Meier , Neoplasias Hepáticas/patologia , Masculino , Camundongos , Microvasos/patologia , Pessoa de Meia-Idade , Invasividade Neoplásica , Fosfoglicerato Quinase/sangue , Valor Preditivo dos Testes , Modelos de Riscos Proporcionais , Proteínas Ribossômicas/genética , Fatores de Risco , Regulação para Cima , alfa-Fetoproteínas/metabolismo
5.
Mol Vis ; 17: 779-91, 2011 Mar 23.
Artigo em Inglês | MEDLINE | ID: mdl-21527995

RESUMO

PURPOSE: To identify candidate protein biomarkers in sera indicative of acute retinal injury. METHODS: We used laser photocoagulation as a model of acute retinal injury in Rhesus macaques. In a paired-control study design, we collected serum from each animal (n=6) at 4 h, 1 day, and 3 days following a mock procedure and then again following retinal laser treatment that produced mild lesions. Samples were fractionated by isoelectric focusing, digested with trypsin, and analyzed by liquid chromatography/tandem mass spectrometry (LC-MS/MS). Spectral counting was used to determine relative protein abundances and identify proteins with statistically significant differences between control and treated sera. RESULTS: Mild retinal injury was confirmed by fundus photography and histological examination. The average number of total proteins detected by LC-MS/MS was 908±82 among samples from all three time points. Following statistical analysis and employing stringent filtering criteria, a total of 19 proteins were identified as being significantly more abundant in sera following laser-induced retinal injury, relative to control sera. Many of the proteins detected were unique to one time point. However, four proteins (phosphoglycerate kinase 1, keratin 18, Lewis alpha-3-fucosyltransferase, and ephrin receptor A2) showed differences that were significant at both 4 h and 1 day after laser treatment, followed by a decrease to baseline levels by day 3. CONCLUSIONS: A serum biomarker response to mild retinal laser injury was demonstrated in a primate model. Among the proteins detected with highest significant differences, most are upregulated within 24 h, and their appearance in the serum is transient. It is conceivable that a panel of these proteins could provide a means for detecting the acute-phase response to retinal injury. Further investigation of these candidate biomarkers and their correlation to retinal damage is warranted.


Assuntos
Traumatismos Oculares/sangue , Fucosiltransferases/sangue , Queratina-18/sangue , Fosfoglicerato Quinase/sangue , Receptores da Família Eph/sangue , Retina/metabolismo , Animais , Biomarcadores/sangue , Cromatografia Líquida , Modelos Animais de Doenças , Traumatismos Oculares/genética , Feminino , Fucosiltransferases/genética , Perfilação da Expressão Gênica , Focalização Isoelétrica , Queratina-18/genética , Fotocoagulação/efeitos adversos , Macaca mulatta , Fosfoglicerato Quinase/genética , Proteômica , Receptores da Família Eph/genética , Retina/lesões , Retina/patologia , Espectrometria de Massas em Tandem , Tripsina/metabolismo
6.
Cancer Biomark ; 7(1): 25-37, 2010.
Artigo em Inglês | MEDLINE | ID: mdl-21045262

RESUMO

The immunogenic nature of cancer can be explored to distinguish pancreatic cancer from related non-cancer conditions. We describe a liquid-based microarray approach followed by statistical analysis and confirmation for discovery of auto-immune biomarkers for pancreatic cancer. Proteins from the Panc-1 pancreatic cancer cell line were fractionated using a 2-D liquid separation method into over 1052 fractions and spotted onto nitrocellulose coated glass slides. The slides were hybridized with 37 pancreatic cancer sera, 24 chronic pancreatitis sera and 23 normal sera to detect elevated levels of reactivity against the proteins in spotted fractions. The response data obtained from protein microarrays was first analyzed by Wilcoxon Rank-Sum Tests to generate two lists of fractions that positively responded to the cancer sera and showed p-values less than 0.02 in the pairwise comparison between cancer specimens and normal and chronic pancreatitis specimens. The top 3 fractions with the lowest correlations were combined in receiver operating characteristic analyses. The area-under-the-curve (AUC) values are 0.813 and 0.792 for cancer vs. normal and cancer vs. pancreatitis respectively. Outlier-Sum statistics were then applied to the microarray data to determine the existence of outliers exclusive in cancer sera. The selected fractions were identified by LC-MS/MS. We further confirmed the occurrence of outliers with three proteins among cancer samples in a confirmation experiment using a separate dataset of 165 serum samples containing 48 cancer sera and 117 non-cancer controls. Phosphoglycerate kinase 1 (PGK1) elicited greater reactivity in 20.9% (10 in 48) of the samples in the cancer group, while no outlier was present in the non-cancer groups.


Assuntos
Anticorpos Antineoplásicos/sangue , Autoanticorpos/sangue , Neoplasias Pancreáticas/imunologia , Idoso , Linhagem Celular Tumoral , Humanos , Pessoa de Meia-Idade , Pâncreas/imunologia , Pancreatite/imunologia , Fosfoglicerato Quinase/sangue , Análise Serial de Proteínas/métodos , Curva ROC
7.
J Immunoassay Immunochem ; 29(3): 220-33, 2008.
Artigo em Inglês | MEDLINE | ID: mdl-18569371

RESUMO

Phosphoglycerate kinase (PGK1) is a key enzyme in glycolysis that can also be released from certain cells. In the extracellular milieu, PGK1 reportedly acts as a disulphide reductase to activate plasmin, resulting in the production of angiostatin, a potent angiogenesis inhibitor. Certain cancer cell lines secrete unusually large amounts of PGK1, raising the possibility that serum PGK1 levels can be used to screen for cancer. To facilitate the characterization of the PGK1 secretory pathway and to monitor serum levels of PGK1, we have developed a sensitive sandwich ELISA using an immuno-affinity-purified chicken polyclonal antibody for capturing PGK1 and an immuno-affinity-purified rabbit polyclonal antibody for detecting it. The assay is about 10-fold more sensitive than other reported PGK1 ELISAs. We used the ELISA to quantify the amount of PGK1 released from HeLa cells and PGK1 serum levels in cancer patients. Of 10 cancer patients whose serum was tested, 3 of 4 with pancreatic cancer had 65-900% higher levels of PGK1 than that found in normal serum.


Assuntos
Biomarcadores Tumorais/sangue , Ensaio de Imunoadsorção Enzimática/métodos , Neoplasias/enzimologia , Fosfoglicerato Quinase/sangue , Fosfoglicerato Quinase/metabolismo , Células HeLa , Humanos , Fosfoglicerato Quinase/imunologia , Proteínas Recombinantes/metabolismo , Reprodutibilidade dos Testes , Sensibilidade e Especificidade
8.
Proteomics ; 6(7): 2259-72, 2006 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-16493704

RESUMO

Pancreatic ductal adenocarcinoma (PDAC) is a common malignancy with a very low 5-year survival rate. Currently, there are no valid markers for early detection and targets for therapy. Here, we used 2-DE to analyze the protein profiles of eight PDAC specimens and paired adjacent nontumor tissues. MS was used to identify 15 protein spots that were found to be overexpressed in PDAC tissues versus adjacent controls. One of them was identified as phosphoglycerate kinase (PGK) 1, a secretable glycolytic enzyme known to participate in angiogenesis. Immunohistochemical analysis of 63 PDAC specimens revealed moderate to strong expression of PGK1 in >70% of the tumors. Further Western blotting analysis of cells from tumor and adjacent nontumor tissues obtained by laser capture microdissection confirmed the enhanced expression of PGK1 in tumor cells. Furthermore, the serum levels of PGK1 were significantly higher in PDAC patients (n = 21) than in the control group (n = 25) (p < 0.005), as determined by ELISA. These observations indicate that protein profile analysis using a combination of 2-DE and MS provides an effective strategy for identifying biomarkers that may have diagnostic potential for PDAC, and identify PGK1 as a potential biomarker and/or therapeutic target for PDAC.


Assuntos
Adenocarcinoma/enzimologia , Carcinoma Ductal Pancreático/enzimologia , Proteínas de Neoplasias/biossíntese , Proteínas de Neoplasias/genética , Neoplasias Pancreáticas/enzimologia , Fosfoglicerato Quinase/biossíntese , Fosfoglicerato Quinase/genética , Proteoma , Adenocarcinoma/patologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Sequência de Aminoácidos , Biomarcadores Tumorais , Western Blotting , Carcinoma Ductal Pancreático/patologia , Feminino , Humanos , Masculino , Microdissecção , Pessoa de Meia-Idade , Dados de Sequência Molecular , Proteínas de Neoplasias/sangue , Neoplasias Pancreáticas/patologia , Fosfoglicerato Quinase/sangue
10.
Eur J Neurol ; 7(1): 111-3, 2000 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-10809925

RESUMO

Phosphoglycerate kinase (PGK) catalyses the transfer of the acylphosphate group of 1,3-diphosphoglycerate to ADP with formation of 3-phosphoglycerate and ATP in the terminal stage of the glycolytic pathway. Two young brothers are presented who both experienced muscle pain, cramps and stiffness shortly after beginning heavy exercise. After these episodes they noticed that the urine was dark brown, indicating rhabdomyolysis and myoglobinuria. The neurological examinations were without remarks. There was no lactate increase in the ischaemic forearm exercise test. Both had very low PGK levels in muscle, erythrocytes, leukocytes and fibroblasts. This is the first family with more than one affected case of PGK deficiency and exercise-induced stiffness, myalgia and rhabdomyolysis. The clinical manifestations may resemble myophosphorylase deficiency (McArdle's disease: glycogenosis Type V) and muscle phosphofructokinase deficiency (Tarui's disease: glycogenosis Type VII). PGK deficiency is inherited as an X-linked trait and may show other features such as mental retardation and/or haemolytic anaemia.


Assuntos
Doença de Depósito de Glicogênio Tipo VII/diagnóstico , Doença de Depósito de Glicogênio Tipo V/diagnóstico , Erros Inatos do Metabolismo/diagnóstico , Erros Inatos do Metabolismo/enzimologia , Fosfoglicerato Quinase/deficiência , Adolescente , Adulto , Diagnóstico Diferencial , Tolerância ao Exercício , Humanos , Masculino , Erros Inatos do Metabolismo/urina , Músculo Esquelético/enzimologia , Mioglobinúria/diagnóstico , Núcleo Familiar , Fosfoglicerato Quinase/sangue
11.
Am J Perinatol ; 14(5): 297-302, 1997 May.
Artigo em Inglês | MEDLINE | ID: mdl-9259948

RESUMO

A case-control study was performed in eight pairs of women to determine whether preeclamptic women developed abnormalities in minor hemoglobins, glycolytic enzymes, or other blood components that might provide insight into the pathophysiology of preeclampsia, or that in combination might be used as a marker for the condition. These variables and standard clinical tests were analyzed as discriminators between preeclamptic and control women. The subjects were matched for age, ethnicity, parity, and gestational age. Blood samples were taken at the time of diagnosis of preeclampsia and at comparable gestational ages for matched normal controls. Variables differing significantly between groups included increases in uric acid (UA), low-density lipoproteins (LDL), phosphoglycerate kinase (PGK), and mean platelet volume (MPV), and decreases in glyceraldehyde phosphate dehydrogenase (G3PD) in preeclamptic women compared to normal controls. Discriminant analysis revealed the following function to separate the groups: 0.7764 (UA) + 0.8086 (PGK) -0.7032 (G3PD) + 0.1399 (LDL) -0.2386 (MPV). A discriminant score of > or = 275 indicated a > or = 90% probability of preeclampsia. The results are consistent with perturbations in red cell glycolysis in preeclampsia. Further prospective studies are warranted to test the efficacy of this discriminant function in predicting preeclampsia.


Assuntos
Eritrócitos/enzimologia , Hemoglobinas/metabolismo , Pré-Eclâmpsia/sangue , Adulto , Biomarcadores/sangue , Estudos de Casos e Controles , Cromatografia por Troca Iônica , Análise Discriminante , Feminino , Idade Gestacional , Gliceraldeído-3-Fosfato Desidrogenases/sangue , Humanos , Recém-Nascido , Lipoproteínas LDL/sangue , Pessoa de Meia-Idade , Fosfoglicerato Quinase/sangue , Pré-Eclâmpsia/diagnóstico , Pré-Eclâmpsia/etiologia , Valor Preditivo dos Testes , Gravidez , Resultado da Gravidez , Ácido Úrico/sangue
12.
Artigo em Inglês | MEDLINE | ID: mdl-9418003

RESUMO

14CO2 production from [1-14C] glucose, the rate of glycolysis measured by the value of lactate production and the activities of various enzymes were determined in buffalo erythrocytes. Buffalo red cell glycolytic metabolites were estimated and used for the calculation of the mass action ratios of reactions catalyzed by the glycolytic enzymes of Bubalus bubalis. A comparison of the values of the mass action ratios with the equilibrium constants of the various glycolytic reactions indicate that hexokinase, phosphofructokinase, phosphoglycerate kinase and pyruvate kinase reactions are displaced from equilibrium, suggesting a regulatory role for each of these enzymes in buffalo erythrocyte glycolysis.


Assuntos
Glicemia/metabolismo , Búfalos/sangue , Eritrócitos/metabolismo , Animais , Dióxido de Carbono/sangue , Glicólise , Hexoquinase/sangue , Cinética , Fosfofrutoquinase-1/sangue , Fosfoglicerato Quinase/sangue , Piruvato Quinase/sangue
13.
Blood ; 88(4): 1479-87, 1996 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-8695869

RESUMO

Rabbit erythrocytes of progressively increasing age were isolated using an avidin-biotin affinity technique and the activity of protein kinases and other enzymes was analyzed in cytosols and membranes from the isolated cells. The activities of cytosolic protein kinase C (PKC), cAMP-dependent kinase (PKA), and casein kinase type I and II (CKI and II) were all found to undergo an age-dependent decrease of twofold to fourfold over the 8-week lifespan of the cells. Membrane-associated tyrosine kinase showed little or no decrease, but membrane-associated CKI showed a dramatic eightfold decrease over the 8-week period. By contrast, various cytosolic enzymes, including lactate dehydrogenase, phosphoglycerate kinase, pyruvate kinase, and acid phosphatase, showed no change in activity over the same time period. Density-separated human erythrocytes showed qualitatively similar decreases in cytosolic protein kinase activities in the densest fractions, which contain the oldest cells. Our results show that aging erythrocytes undergo progressive loss of protein kinases that may adversely affect various cellular processes. The age-dependent loss of kinase activity reported here is one of the most striking manifestations of erythrocyte senescence yet to be reported.


Assuntos
Envelhecimento Eritrocítico , Eritrócitos/enzimologia , Proteína Quinase C/sangue , Fosfatase Ácida/sangue , Animais , Citosol/enzimologia , Membrana Eritrocítica/enzimologia , Humanos , L-Lactato Desidrogenase/sangue , Fosfoglicerato Quinase/sangue , Fosfoproteínas/sangue , Piruvato Quinase/sangue , Coelhos
14.
Blood Cells Mol Dis ; 21(3): 179-81, 1995.
Artigo em Inglês | MEDLINE | ID: mdl-8673469

RESUMO

The molecular abnormality of a phosphoglycerate kinase variant associated with severe red cell enzyme deficiency ( about 4% of normal) and episodes of hemolysis with jaundice was examined. The Michaelis constants for the substrates and co-enzymes (1.3-diphosphoglycerate, 3-phosphoglycerate, ATP and ADP) were not grossly different from that of normal. However that variant enzyme was very labile in vitro. Nucleotide sequence analysis of the variant cDNA revealed a deletion of codon AAG in exon 7. The codon deletion should result in the election of one of the tandem lysine residues existing at amino acid 190-191 of the enzyme protein. Based on the three dimensional structure of the protein, molecular instability could could be induced by the deletion of a lysine residue.


Assuntos
Anemia Hemolítica Congênita não Esferocítica/genética , Variação Genética , Fosfoglicerato Quinase/genética , Adulto , Anemia Hemolítica Congênita não Esferocítica/enzimologia , Sequência de Bases , Análise Mutacional de DNA , Eritrócitos/enzimologia , Humanos , Cinética , Masculino , Dados de Sequência Molecular , Fosfoglicerato Quinase/sangue , Fosfoglicerato Quinase/química , Fosfoglicerato Quinase/deficiência , Conformação Proteica , Deleção de Sequência
15.
Exp Parasitol ; 75(1): 10-8, 1992 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-1639156

RESUMO

Multiplication of the human malaria parasite Plasmodium falciparum within red blood cells is an energy-dependent process and glucose consumption increases dramatically in infected red blood cells (IRBC) versus normal red blood cells (NRBC). The major pathway for glucose metabolism in P. falciparum IRBC is anaerobic glycolysis. Phosphoglycerate kinase (PGK) is one of the key enzymes of this pathway as it generates ATP. We found that the PGK specific activity in P. falciparum IRBC is seven times higher than that in NRBC. The parasitic origin of the increase in PGK activity is confirmed by isoelectric focusing. Indeed, two P. falciparum isoenzymes with neutral isoelectric points were detected. P. falciparum PGK in purified form has a molecular mass of 48 kDa. Antiserum raised against purified P. falciparum PGK specifically recognizes the 48-kDa protein band in P. falciparum and also reacts with P. berghei and P. yoelii IRBC lysates but does not cross-react with PGK associated with NRBC.


Assuntos
Eritrócitos/enzimologia , Malária Falciparum/enzimologia , Fosfoglicerato Quinase/isolamento & purificação , Plasmodium falciparum/enzimologia , Animais , Western Blotting , Cromatografia por Troca Iônica , Eritrócitos/parasitologia , Humanos , Soros Imunes , Ponto Isoelétrico , Cinética , Camundongos , Camundongos Endogâmicos BALB C , Peso Molecular , Fosfoglicerato Quinase/sangue , Fosfoglicerato Quinase/metabolismo
16.
Am J Med Genet ; 42(5): 660-4, 1992 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-1632433

RESUMO

A 2 8/12-year-old boy with severe growth failure and mental retardation was found to have a maternally derived tandem duplication of the long arm of X chromosome, dup(X) (q13.3----q21.2). Karyotypic interpretation was further confirmed in this patient by a double gene dose for red blood cell phosphoglycerate kinase. DNA replication study showed that the duplicated X chromosome was always late replicating in peripheral blood lymphocytes as well as in skin fibroblasts from the mother. Endocrine studies in the patient demonstrated growth hormone deficiency. Magnetic resonance imaging of the head then disclosed the empty sella syndrome. This appears to be the first report of a dup(Xq) patient associated with a growth hormone deficiency and the empty sella syndrome. We emphasize that duplication of the proximal Xq in males represents another microduplication syndrome (Thode-Leonard syndrome).


Assuntos
Síndrome da Sela Vazia/genética , Hormônio do Crescimento/deficiência , Família Multigênica/genética , Aberrações dos Cromossomos Sexuais/genética , Cromossomo X , Pré-Escolar , Feminino , Humanos , Cariotipagem , Masculino , Fosfoglicerato Quinase/sangue , Fosfoglicerato Quinase/genética
17.
J Gerontol ; 46(6): B213-6, 1991 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-1940071

RESUMO

We investigated whether age-dependent reactivation of a repressed X-linked gene occurs. Subjects were female mice, carrying the X-autosomal translocation, T(X;16)16H (Searle's translocation). The mice were also heterozygous, for the X-linked gene coding for phosphoglycerate kinase (T16H pgk-1b/+ pgk-1a and pgk-1a was selectively repressed in these mice (McMahon and Monk, 1983). An electrophoretic method was applied to determine the PGK-1 allozyme patterns in blood, bone marrow, brain, gastrointestinal tract, liver, heart, spleen, and uterus (including tumor tissues when found). Samples were collected from mice of three different ages: 2 months (n = 4), 11 to 12 months (n = 10), or 18 to 21 months (n = 15). The lowest detection limit of the relative cellular population expressing the PGK-1A allozyme was found to be 2%, which was sensitive enough to detect the reported reactivation ratio (more than 10% of cells in a lower power microscopic field). We could not detect PGK-1A activity in any organ, including tumors in any age group, leading to the conclusion that reactivation of the repressed pgk-1a gene did not occur during aging.


Assuntos
Envelhecimento/genética , Regulação Enzimológica da Expressão Gênica/fisiologia , Ligação Genética/genética , Fosfoglicerato Quinase/genética , Translocação Genética/genética , Cromossomo X , Animais , Encéfalo/enzimologia , Eletroforese , Eritrócitos/enzimologia , Feminino , Genótipo , Fígado/enzimologia , Camundongos , Camundongos Endogâmicos C3H , Camundongos Endogâmicos , Miocárdio/enzimologia , Neoplasias Experimentais/enzimologia , Fenótipo , Fosfoglicerato Quinase/análise , Fosfoglicerato Quinase/sangue , Fosfoglicerato Quinase/classificação , Baço/enzimologia , Útero/enzimologia
18.
Lab Delo ; (5): 41-3, 1991.
Artigo em Russo | MEDLINE | ID: mdl-1715942

RESUMO

The developed modification may be used in clinical practice as a diagnostic test and in mass screenings for phosphoglycerate kinase deficiency: more than a hundred tests may be made in a day. If blood samples have to be sent by mail, their quality is unchanged for up to 7 days.


Assuntos
Eritrócitos/enzimologia , Programas de Rastreamento/métodos , Fosfoglicerato Quinase/deficiência , Criança , Humanos , Fosfoglicerato Quinase/sangue , Coloração e Rotulagem
19.
Indian J Med Res ; 92: 21-3, 1990 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-2189828

RESUMO

Red cell phosphoglycerate kinase (PGK) activity was determined in normal individuals and patients with, type I (insulin-dependent) diabetes and insulin treated diabetes. The PGK activity was significantly (P less than 0.001) elevated in diabetes, however it is restored to normalcy after insulin treatment (normal 282.54 +/- 9.46, type I diabetic 342.06 +/- 6.24, insulin treated diabetic 292.66 +/- 7.12 IU/g haemoglobin at 37 degrees C). No significant alteration was observed in the percentage of PGK bound to the membrane fraction of red cells in all the three conditions. The results indicate that the increased PGK activity is a result of a regulatory mechanism induced by the fluctuation of ATP level in response to elevated Na:K pump rate of erythrocytes in type I diabetes mellitus.


Assuntos
Diabetes Mellitus Tipo 1/enzimologia , Eritrócitos/enzimologia , Insulina/uso terapêutico , Fosfoglicerato Quinase/sangue , Diabetes Mellitus Tipo 1/tratamento farmacológico , Humanos
20.
Comp Biochem Physiol B ; 97(1): 47-8, 1990.
Artigo em Inglês | MEDLINE | ID: mdl-2147643

RESUMO

1. The activity of 21 red cell enzymes and three red cell metabolic intermediates were measured in adult Dasyurus viverrinus and compared with published data on other marsupials. 2. Phosphofructokinase (PFK), glyceraldehyde dehydrogenase (GAPD) and phosphoglycerate kinase (PGK) were elevated in comparison to other marsupials. 3. Enolase (ENO) and 2,3-diphosphoglycerate (2,3 DPG) were lower than in other marsupials.


Assuntos
Eritrócitos/enzimologia , Gliceraldeído-3-Fosfato Desidrogenases/sangue , Marsupiais/sangue , Fosfofrutoquinase-1/sangue , Fosfoglicerato Quinase/sangue , Animais , Eritrócitos/metabolismo , Feminino , Masculino
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