RESUMO
A distinct cluster of highly pathogenic avian influenzaviruses of subtype A(H5N1) has been found to emergewithin clade 2.2.1.2 in poultry in Egypt since summer2014 and appears to have quickly become predominant.Viruses of this cluster may be associated withincreased incidence of human influenza A(H5N1) infectionsin Egypt over the last months.
Assuntos
Virus da Influenza A Subtipo H5N1/genética , Influenza Aviária/virologia , Influenza Humana/virologia , Aves Domésticas , Animais , Doenças Transmissíveis Emergentes , Egito/epidemiologia , Glicoproteínas de Hemaglutininação de Vírus da Influenza/genética , Humanos , Virus da Influenza A Subtipo H5N1/isolamento & purificação , Influenza Aviária/diagnóstico , Influenza Humana/epidemiologia , Filogenia , Aves Domésticas/virologia , Doenças das Aves Domésticas/epidemiologia , RNA Viral/genética , Análise de Sequência de RNARESUMO
Twin pregnancy in mares is one of the leading causes of abortions. Abortion invariably impacts both fetuses. This report describes an unusual case of a twin surviving to term following the abortion of its co-twin at 9 months in a 7-year-old Egyptian Arabian mare. At the time of abortion at 9 months of gestation, the size of the aborted fetus was equivalent to one of approximately 5 months of age while the age of the live co-twin was 9 months. Both fetuses were males. A skin sample was collected from the aborted fetus and hair samples were collected from the dam, sire and live foal for parentage analysis. The parentage analysis confirmed that both fetuses were by the same dam and sire stallion. The authors suggest several scenarios to explain this condition. This report describes a unique case of a twin surviving to term following the abortion of its co-twin at 9 months in a mare.
Assuntos
Aborto Animal , Cavalos , Feminino , Animais , Gravidez , Masculino , Gêmeos , Gravidez de GêmeosRESUMO
Quiescent mouse NIH 3T3 cells responded to microinjection of activated ras p21 with a rapid and sustained rise in intracellular pH (approximately 0.17 pH units). The p21-induced pH change was inhibited by amiloride treatment or growth of cells in medium low in sodium, suggesting a role for the Na+/H+ antiporter. Amiloride was found to suppress p21-induced mitosis, also.
Assuntos
Concentração de Íons de Hidrogênio , Proteínas Proto-Oncogênicas/farmacologia , Amilorida/farmacologia , Animais , Proteínas de Transporte/fisiologia , Linhagem Celular , DNA/biossíntese , Replicação do DNA/efeitos dos fármacos , Camundongos , Microinjeções , Microscopia de Fluorescência , Proteínas Proto-Oncogênicas p21(ras) , Ratos , Trocadores de Sódio-HidrogênioRESUMO
The role of epidermal growth factor in the regulation of the proteolytic and RBC cytolytic activity of the A431 cancer cell line has been evaluated using our previously described gelatin/polyacrylamide electrophoretic assay and tumor-induced RBC cytolysis assay, respectively. A431 cells maintained in 10% fetal bovine serum were actively cytolytic for RBC (release index, 53.0 +/- 2.9%), whereas serum-starved cells maintained in serum-free medium were not cytolytic for RBC. RBC cytotoxicity was restored by adding as little as 3.4 pM epidermal growth factor to the serum-deprived cells. The RBC cytolytic stimulating activity of epidermal growth factor could be mimicked by the metal chelating agent 1,10-phenanthroline, suggesting a possible role for calcium ions in the action of epidermal growth factor and proteases. An enriched cell membrane preparation of A431 cells was also cytolytic for RBC but was unaffected by metal chelating agents. RBC-induced cytotoxicity was inhibited by the protease inhibitor leupeptin. Gelatin substrate gels of enriched A431 cell membrane preparations and serum-free supernatants revealed a pattern of high- and low-molecular-weight proteases that were stimulated by metal chelators and inhibited by leupeptin. The activity of these proteases appears to be regulated by epidermal growth factor by a process that may involve divalent cations.
Assuntos
Carcinoma de Células Escamosas/enzimologia , Fator de Crescimento Epidérmico/farmacologia , Eritrócitos/efeitos dos fármacos , Peptídeo Hidrolases/análise , Carcinoma de Células Escamosas/fisiopatologia , Ácido Edético/farmacologia , Eletroforese em Gel de Poliacrilamida , Receptores ErbB/fisiologia , Eritrócitos/patologia , Humanos , Peso Molecular , Biossíntese de Proteínas , Tosilina Clorometil Cetona/farmacologia , Células Tumorais CultivadasRESUMO
Peripheral T-cell lymphomas (PTCLs) are aggressive lymphomas with no effective upfront standard treatment and ineffective options in relapsed disease, resulting in poorer clinical outcomes as compared with B-cell lymphomas. The adoptive transfer of T cells engineered to express chimeric antigen receptors (CARs) is a promising new approach for treatment of hematological malignancies. However, preclinical reports of targeting T-cell lymphoma with CARs are almost non-existent. Here we have designed a CAR, CD4CAR, which redirects the antigen specificity of CD8+ cytotoxic T cells to CD4-expressing cells. CD4CAR T cells derived from human peripheral blood mononuclear cells and cord blood effectively redirected T-cell specificity against CD4+ cells in vitro. CD4CAR T cells efficiently eliminated a CD4+ leukemic cell line and primary CD4+ PTCL patient samples in co-culture assays. Notably, CD4CAR T cells maintained a central memory stem cell-like phenotype (CD8+CD45RO+CD62L+) under standard culture conditions. Furthermore, in aggressive orthotropic T-cell lymphoma models, CD4CAR T cells efficiently suppressed the growth of lymphoma cells while also significantly prolonging mouse survival. Combined, these studies demonstrate that CD4CAR-expressing CD8+ T cells are efficacious in ablating malignant CD4+ populations, with potential use as a bridge to transplant or stand-alone therapy for the treatment of PTCLs.
Assuntos
Transferência Adotiva , Linfócitos T CD8-Positivos/imunologia , Linfoma de Células T Periférico/terapia , Proteínas Mutantes Quiméricas/genética , Receptores Artificiais/genética , Animais , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD4-Positivos/patologia , Linfócitos T CD8-Positivos/citologia , Linfócitos T CD8-Positivos/transplante , Engenharia Celular , Linhagem Celular Tumoral , Técnicas de Cocultura , Citotoxicidade Imunológica , Expressão Gênica , Humanos , Leucemia/genética , Leucemia/imunologia , Leucemia/patologia , Linfoma de Células T Periférico/genética , Linfoma de Células T Periférico/imunologia , Linfoma de Células T Periférico/mortalidade , Masculino , Camundongos , Camundongos Endogâmicos NOD , Proteínas Mutantes Quiméricas/imunologia , Transplante de Neoplasias , Cultura Primária de Células , Receptores Artificiais/imunologia , Análise de SobrevidaRESUMO
Highly pathogenic avian influenza (HPAI) viruses of the H5N1 subtype are enzootic in poultry populations in different parts of the world, and have caused numerous human infections in recent years, particularly in Egypt. However, no sustained human-to-human transmission of these viruses has yet been reported. We tested nine naturally occurring Egyptian H5N1 viruses (isolated in 2014-2015) in ferrets and found that three of them transmitted via respiratory droplets, causing a fatal infection in one of the exposed animals. All isolates were sensitive to neuraminidase inhibitors. However, these viruses were not transmitted via respiratory droplets in three additional transmission experiments in ferrets. Currently, we do not know if the efficiency of transmission is very low or if subtle differences in experimental parameters contributed to these inconsistent results. Nonetheless, our findings heighten concern regarding the pandemic potential of recent Egyptian H5N1 influenza viruses.
Assuntos
Virus da Influenza A Subtipo H5N1/genética , Infecções por Orthomyxoviridae/epidemiologia , Infecções por Orthomyxoviridae/veterinária , Animais , Antivirais/farmacologia , Bioensaio , Cães , Egito/epidemiologia , Inibidores Enzimáticos/farmacologia , Furões , Expressão Gênica , Células HeLa , Humanos , Virus da Influenza A Subtipo H5N1/classificação , Virus da Influenza A Subtipo H5N1/efeitos dos fármacos , Virus da Influenza A Subtipo H5N1/isolamento & purificação , Células Madin Darby de Rim Canino , Neuraminidase/antagonistas & inibidores , Neuraminidase/genética , Neuraminidase/metabolismo , Infecções por Orthomyxoviridae/tratamento farmacológico , Infecções por Orthomyxoviridae/transmissão , Filogenia , Medição de Risco , Carga Viral/efeitos dos fármacos , Proteínas Virais/antagonistas & inibidores , Proteínas Virais/genética , Proteínas Virais/metabolismoRESUMO
The modulation of responsive genes by hormonal stimulation is an attractive in vitro model system for the study of a wide variety of biological processes. Using this methodology we have investigated the effect of the human oncogene protein p21ras on mitosis using mouse mammary tumor virus long terminal repeat (MMTV-LTR)-directed gene expression. Following the induction of p21 protein, abnormal mitotic figures were scored in metaphase and anaphase. Elevated expression of p21 was associated with marked increase in the proportion of abnormal mitoses most significantly during the metaphase. Concomitant with a three fold increase in p21 levels, abnormal mitosis rose from 14.0% to 27.25%. The increase in abnormal mitosis corresponded to a 225% increase in abnormal metaphase. The p21-induced mitotic abnormalities were exhibited as lagging chromosomes in prometaphase, 3 group metaphase and C-metaphase. In addition, high expression of p21 was accompanied by significant changes in the cell morphology and fine ultrastructure, e.g. disorganization of actin, the extensive formation of microvilli on the plasma membrane and marked dilatation of the rough endoplasmic reticulum. The mitotic and structural changes were reversible upon removal of dexamethasone and decline of p21 production to its basal levels. Our results identify an important biological effect of ras p21 during mitosis and the early stages of neoplastic transformation.
Assuntos
Mitose , Proteínas Proto-Oncogênicas p21(ras)/genética , Animais , Linhagem Celular , Cromossomos/ultraestrutura , Células Clonais , Imunofluorescência , Vírus do Tumor Mamário do Camundongo/genética , Camundongos , Camundongos Endogâmicos , Sequências Repetitivas de Ácido Nucleico , TransfecçãoRESUMO
We describe a simple procedure to measure precisely the tip diameter of glass micropipets. This procedure can improve the precision of microinjection and some electrophysiological measurements due to their extreme sensitivity to the pipet's exact geometric dimensions. The technique is nondestructive and readily carried out as a brief, extra step in these applications. Digital measurements of the threshold pressures for gas bubbling from (gas filled) pipets submerged in methanol were used to calculate the corresponding inner tip diameters using the LaPlace equation. Direct measurements of the inner tip diameters (from 0.2-5.0 mu) using scanning electron microscopy confirmed the validity of this indirect, simple procedure.
Assuntos
Eletrofisiologia/instrumentação , Microinjeções/instrumentaçãoRESUMO
Ewing's sarcoma (ES) and related neoplasias are characterized by the reciprocal translocation, t(11;22)(q24;q12). The translocation has been reported to generate a fusion gene between the EWS (a previously undescribed gene on chromosome 22) and FLI1 genes. We report a similar translocation of EWS and FLI1 in an Askin's tumor cell line (SK-NM-C). Further, we describe an alternative translocation in an ES cell line (#5838) in which the 5' end of the EWS gene is juxtaposed to the 3' end of the ERG gene. The ERG gene is on chromosome 21, but no microscopically visible changes in chromosome 21 were observed. Elevated steady state levels of the EWS/ERG fusion gene transcript were detected in the #5838 cell line. This is the first report of a structural alteration of ERG in human cancer. Also, it confirms a general mechanism of generating putative oncogenic fusion genes by placing an ETS DNA binding domain in direct proximity to the carboxy terminus domain (CTD) related region of the EWS gene.
Assuntos
Sarcoma de Ewing/genética , Translocação Genética , Sequência de Bases , Clonagem Molecular , Humanos , Hibridização in Situ Fluorescente , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Células Tumorais CultivadasRESUMO
The biological effects of EGF and PDGF growth factors on A172 and hEGFr-3T3 cell lines were studied using RBC induced cytolysis and polyacrylamide-gelatin gel electrophoresis assays. The authors report that growth factor-induced cytotoxicity in these cells is mediated by proteolytic enzymes. Treatment of A172 cells with either EGF or PDGF resulted in marked increase of their cytotoxicity (Release Index = 150%). Similarly, RBC induced release index by hEGFr-3T3 cells was elevated to 420% in the presence of 3.4 pM of EGF. However, in A172 cells, PDGF did not have a significant effect on DNA and protein synthesis indicating that stimulation of proteolytic activity is independent of the growth factor signaling pathway. Growth factor induced cytotoxicity was significantly reduced by protease inhibitors in both cell lines. Using EDTA and leupeptin several proteolytic species were identified and localized to cellular membranes as evidenced by polyacrylamide-gelatin electrophoresis assay. These data suggest that growth factors regulate the activation or secretion of proteolytic enzymes in cancer cells and may mediate the invasive and metastatic behavior of these cells.
Assuntos
Fator de Crescimento Epidérmico/farmacologia , Peptídeo Hidrolases/metabolismo , Fator de Crescimento Derivado de Plaquetas/farmacologia , Células Tumorais Cultivadas/efeitos dos fármacos , Amilorida/farmacologia , Animais , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Cicloeximida/farmacologia , Ativação Enzimática , Hemólise/efeitos dos fármacos , Humanos , Leupeptinas/farmacologia , Camundongos , Tosilina Clorometil Cetona/farmacologia , Células Tumorais Cultivadas/enzimologiaRESUMO
The protein products (p21) of the ras cellular proto-oncogenes are thought to transduce membrane signals necessary for the induction of cell division. However, there is uncertainty as to the precise role of ras p21 in mediating ligand-membrane receptor signals leading to cell differentiation. Treatment of rat phaeochromocytoma cells (PC12) with nerve growth factor (NGF) results in the induction of a number of phenotypic characteristics of sympathetic neurones, including cessation of cell division and outgrowth of neuronal processes (neurites). Here we report that microinjection of antibody to ras p21 into PC12 cells inhibited neurite formation and resulted in temporary regression of partially extended neurites, an effect which was observed up to 36 h after initiation of NGF treatment. Neurite formation induced by cyclic AMP was unaffected by injection of anti-p21 antibody. These results indicate that p21 is involved in the initiation phase of NGF-induced neurite formation in PC12 cells and has a role in hormone-mediated cellular responses distinct from cell proliferation.
Assuntos
Diferenciação Celular/efeitos dos fármacos , Fatores de Crescimento Neural/farmacologia , Proteínas Proto-Oncogênicas/fisiologia , Animais , Anticorpos/imunologia , Linhagem Celular , AMP Cíclico/fisiologia , Microinjeções , Feocromocitoma/patologia , Proteínas Proto-Oncogênicas/imunologia , Ratos , Fatores de TempoRESUMO
The human oncogene ras p21 was transfected into human lung fibroblast WI-38 diploid cells. All of the clones that were isolated (n = 36), exhibited rapid growth and transformed morphology which was ascertained by both transmission and scanning electron microscopy as extensive formations of microvilli on the plasma membrane, marked distortion of the nuclear membrane and increased number of pinocytotic vesicles in the cortical cytoplasm. The frequency of abnormal metaphases rose from 15% in parental WI-38 cells to 35.5-48.4% in all ten clones examined. Lagging chromosomes in prometaphase represented 42.1-69.4% of the total abnormal mitoses followed in frequency by 3-group metaphase and C-metaphase. All transformed cells were aneuploid. These data provide evidence for the association between cellular transformation with oncogenic ras and elevated abnormal mitoses in human cells.
Assuntos
Fibroblastos/ultraestrutura , Genes ras , Mitose , Aneuploidia , Linhagem Celular Transformada , Aberrações Cromossômicas , Dexametasona/farmacologia , Diploide , Feminino , Fibroblastos/efeitos dos fármacos , Humanos , Cariotipagem , Pulmão , Microscopia Eletrônica , Proteínas Proto-Oncogênicas p21(ras)/biossíntese , Proteínas Proto-Oncogênicas p21(ras)/fisiologia , Proteínas Recombinantes de Fusão/biossíntese , TransfecçãoRESUMO
A method that permits the in vitro cloning and identification of proteolytic enzyme genes from cDNA expression libraries is described. The method can detect positive proteinase genes within 30 min following the transfer of plaques to nitrocellulose membrane filters. The method is based on the functional expression of fusion lac Z-proteinase protein in lambda gt11 infected Y1090 bacteria and does not require prior knowledge of either the sequence of the cDNA insert or a monoclonal antibody to its encoded antigen. This strategy when coupled with polymerase chain reaction of the cDNA insert using lac Z primer sequences that are flanking the EcoR1 cloning site in gt11 phage permits direct sequencing of the amplified DNA. With this method we have isolated 10 genes expressing protease activity in the human small-cell carcinoma of the lung. The same procedure could be applied to isolate unknown proteinases from cDNA libraries of virtually any eukaryotic cell.
Assuntos
Carcinoma de Células Pequenas/genética , Clonagem Molecular/métodos , Endopeptidases/genética , Sequência de Bases , Carcinoma de Células Pequenas/enzimologia , Eletroforese , Precursores Enzimáticos , Expressão Gênica , Biblioteca Gênica , Genes , Humanos , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Células Tumorais CultivadasRESUMO
1. A new population of neurons with transient expression of NPY immunoreactivity was described in the developing hamster paraventricular thalamic area. The present study was performed to discover whether this phenomenon is due to programmed cell death or apoptosis. 2. Toward this aim, immunocytochemical and electron microscopic examination of the paraventricular thalamic region, as well as DNA electrophoresis of tissue extracted from the described area, was performed on different stages of embryonic and postnatal development. 3. A sudden increase in neuropeptide Y immunoreactivity (NPY-IR) in the paraventricular thalamic area at embryonic day 14 (E14) was the first symptom of neuronal degeneration. 4. Electron microscopy revealed many neurons with large masses of condensed chromatin within nuclei and extracellular bodies. The affected cells had a convoluted shape and condensed cytoplasm. 5. DNA electrophoresis revealed a ladder of bands between 150 and 1000 bp that is specific for internucleosomal DNA fragmentation. 6. The data strongly suggest that developmental disappearance of NPY-IR neurons within the hamster dorsal thalamic area is due to apoptosis.
Assuntos
Apoptose , Degeneração Neural , Neuropeptídeo Y/biossíntese , Tálamo/metabolismo , Animais , Cricetinae , Fragmentação do DNA , Idade Gestacional , Mesocricetus , Microscopia Eletrônica , Neurônios/química , Neurônios/ultraestrutura , Neuropeptídeo Y/genética , Tálamo/embriologia , Tálamo/ultraestruturaRESUMO
Reaction of p-nitrophenyl anthranilate with human serum albumin at pH 8.0 results in esterification of a single anthraniloyl moiety with the hydroxyl group of tyrosine-411. The absorption spectrum of the anthraniloyl group overlaps the fluorescence emission of the single tryptophan residue at position 214. This study complements that of the preceding paper [Suzukida, M., Le, H. P., Shahid, F., McPherson, R. A., Birnbaum, E.R., & Darnall, D. W. (1983) Biochemistry (preceding paper in this issue)] where an azomercurial group was introduced at cysteine-34. Anthraniloyl fluorescence was also quenched by the azomercurial absorption at Cys-34. Thus measurement of resonance energy transfer between these three sites allowed distances to be measured between Cys-34 in domain I, Trp-214 in domain II, and Tyr-411 in domain III of human serum albumin. At pH 7.4 in 0.1 M phosphate the Trp-214 leads to Tyr-411, Tyr-411 leads to Cys-34, and Trp-214 leads to Cys-34 distances were found to be 25.2 +/- 0.6, 25.2 +/- 2.1, and 31.8 +/- 0.8 A, respectively.
Assuntos
Cisteína , Albumina Sérica , Triptofano , Tirosina , Dicroísmo Circular , Transferência de Energia , Humanos , Concentração de Íons de Hidrogênio , Cinética , Inibidores de Proteases , Conformação Proteica , Espectrofotometria , ortoaminobenzoatosRESUMO
We used RNAase protection and restriction fragment length polymorphism assays to detect activating mutations of c-src in a spectrum of human tumours. No mutations were detected at codons 98, 381, 444, and 530. We conclude that mutational activation is not the mechanism of enhancement of pp60c-src-specific kinase activity found in a number of human cancer types.
Assuntos
Genes src , Mutação , Neoplasias/genética , Proteínas Proto-Oncogênicas pp60(c-src)/metabolismo , Sequência de Bases , Códon , Humanos , Dados de Sequência Molecular , Mutagênese Sítio-Dirigida , Neoplasias/enzimologia , Oligodesoxirribonucleotídeos , Polimorfismo de Fragmento de Restrição , Proteínas Tirosina Quinases/genética , Proteínas Tirosina Quinases/metabolismo , Proteínas Proto-Oncogênicas pp60(c-src)/genética , RNA Mensageiro/genéticaRESUMO
To evaluate the proteolytic activities of oncogene-transfected 3T3 cells, we have developed a copolymerized substrate electrophoretic assay that permits the detection of picogram quantities of proteases produced by cells in culture. Our assay involves a gelatin substrate copolymerized in a polyacrylamide gel. Purified cell membrane preparations were run on gels and various protease activities were detected by amido black. Ras-transfected 3T3 cells appear to have a soluble metalloprotease that may be transiently membrane bound and responsible for destruction of red blood cells (RBC). Oncogene-transfected NIH-3T3 cells have been demonstrated to have RBC cytolytic activity. We have previously shown that v-src-transfected 3T3 cells and their cell membranes cause RBC cytolysis which is inhibited by the protease inhibitor leupeptin. Here we report that both H-ras- and K-ras-transfected 3T3 cells and their cell membranes are cytolytic for RBC, but are inhibited by the metalloprotease inhibitor ethylene diamine tetraacetic acid. Using the gelatin substrate gel assay, we determined that some of the proteases were intrinsic to the oncogene expressing cells, while other proteases were secreted into the culture growth medium.
Assuntos
Genes ras , Oncogenes , Peptídeo Hidrolases/genética , Linhagem Celular Transformada , Meios de Cultura , Ácido Edético/farmacologia , Eletroforese em Gel de Poliacrilamida , Eritrócitos , Hemólise/efeitos dos fármacos , Proteínas de Membrana/análise , Peptídeo Hidrolases/análise , Inibidores de Proteases/farmacologia , Dodecilsulfato de Sódio , EspectrofotometriaRESUMO
A cAMP receptor protein (CRP) species was purified from the luminous Vibrio harveyi cells to apparent homogeneity. This protein had a dimeric structure with a molecular weight of 23,000 per subunit. Among all eight nucleotides tested, only cAMP (Kd = 3 to 4 microM at 0 degrees C and 52 microM at 23 degrees C) and cGMP (Kd = 6 to 10 microM at 0 degrees C and 67 microM at 23 degrees C) bound to this protein. Its binding to poly(dI-dC), poly(dA-dT), and DNA fragments isolated from V. harveyi cells were all significantly enhanced by the addition of cAMP. Based on patterns of limited proteolysis by trypsin, this CRP assumes different conformations in the absence and presence of cAMP. Also consistent with this conclusion is the finding that the binding of cAMP to CRP induced about 50% quenching of the CRP fluorescence with a concomitant 3-nm blue shift from the original 336-nm emission peak. The binding of cGMP resulted in similar fluorescence changes but had no apparent effect on the pattern of proteolysis by trypsin. Using an in vitro transcription system known to be dependent on cAMP and Escherichia coli CRP, the synthesis of a run-off transcript product was also significantly enhanced by cAMP and this V. harveyi CRP.
Assuntos
Receptores de AMP Cíclico/isolamento & purificação , Vibrio/análise , AMP Cíclico/metabolismo , Escherichia coli/análise , Conformação Proteica , Receptores de AMP Cíclico/metabolismo , Transcrição Gênica , TrítioRESUMO
A simple and rapid procedure to isolate clones carrying sequences from a specific region of the polytene chromosome of Drosophila is demonstrated. The procedure involves microdissection of the region of interest, amplification of the DNA by PCR using a primer designed to prime the synthesis nonspecifically, labeling of the amplified DNA using the random primer method, and screening of a standard library with the probe to identify and isolate clones carrying sequences homologous to the dissected region. This procedure has the potential to replace the difficult procedure of microcloning, as well as facilitate chromosome walking.
Assuntos
Cromossomos/análise , Clonagem Molecular , DNA/genética , Drosophila melanogaster/genética , Amplificação de Genes , Reação em Cadeia da Polimerase , Animais , Sequência de Bases , Biotina , DNA/isolamento & purificação , Sondas de DNA , Dados de Sequência Molecular , Hibridização de Ácido NucleicoRESUMO
Reaction of human serum albumin with p-nitrophenylanthranilate results in transesterification of the anthraniloyl group to tyrosine 411. Titration of anthraniloyl-Tyr-411-albumin with long chain or short chain fatty acids produces marked changes in the absorption and fluorescence spectra of the anthraniloyl moiety as fatty acids bind in the channel near it. It appears that the anthraniloyl group is a very sensitive probe that can follow binding of small molecules at the 3-AB subdomain of human serum albumin.