RESUMO
The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic has become a major public health issue worldwide. Developing and evaluating rapid and easy-to-perform diagnostic tests is a high priority. The current study was designed to assess the diagnostic performance of an antigen-based rapid detection test (COVID-VIRO®) in a real-life setting. Two nasopharyngeal specimens of symptomatic or asymptomatic adult patients hospitalized in the Infectious Diseases Department or voluntarily accessing the COVID-19 Screening Department of the Regional Hospital of Orléans, France, were concurrently collected. The diagnostic specificity and sensitivity of COVID VIRO® results were compared to those of real-time reverse-transcriptase quantitative polymerase chain reaction (RT-qPCR) results. A subset of patients underwent an additional oropharyngeal and/or saliva swab for rapid testing. A total of 121 patients confirmed to be infected and 127 patients having no evidence of recent or ongoing infection were enrolled for a total of 248 nasopharyngeal swab specimens. Overall, the COVID-VIRO® sensitivity was 96.7% (CI, 93.5%-99.9%). In asymptomatic patients, symptomatic patients having symptoms for more than 4 days and those with an RT-qPCR cycle threshold value ≥ 32, the sensitivities were 100%, 95.8%, and 91.9%, respectively. The concordance between RT-qPCR and COVID VIRO® rapid test results was 100% for the 127 patients with no SARS-CoV-2 infection. The COVID-VIRO® test had 100% specificity and sensitivity greater than 95%, which are better than the recommendations set forth by the WHO (specificity ≥ 97%-100%, sensitivity ≥ 80%). These rapid tests may be particularly useful for large-scale screening in emergency departments, low-resource settings, and airports.
Assuntos
Antígenos Virais/isolamento & purificação , Teste para COVID-19/métodos , COVID-19/diagnóstico , Nasofaringe/virologia , SARS-CoV-2/isolamento & purificação , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Testes Imediatos , Sensibilidade e Especificidade , Adulto JovemRESUMO
Detonation nanodiamonds exhibit strong nonlinear optical properties depending on their electronic properties. In the present paper, the nanodiamond functional groups are chemically modified to obtain nanodiamonds with primary amines on their surface. The optical properties of such nanodiamonds placed in water suspensions are studied and compared with the one of classical detonation nanodiamonds. Transmission, scattering and Z-scan experiments are performed for two different wavelengths (532 nm and 1064 nm). A lower threshold for optical limiting associated to more pronounce non-linear optical effects is detected at the wavelength of 1064 nm compared to the one at 532 nm. This effect may be due to a stronger nonlinear backscattering behavior at 1064 nm. Moreover, a striking result obtained from the Z-scan experiments reveals a completely different behavior of the functionalized nanodiamonds for both wavelengths. This result is discussed in regard to the electronic properties of the material and possible charge transfer.
RESUMO
Nucleocytoplasmic transport of rat liver mRNA is thought to be regulated by a nucleoside triphosphatase whose activity in the intact nuclear envelope is stimulated by the 3'poly(A) tail of poly(A)+ mRNA. In contrast to the liver mRNA, the mRNA from rat brain contains a great population of poly(A)- mRNA's that does not appear until after birth. Measurements of the nuclear-envelope-associated enzyme activities involved in mRNA transport, and their dependence on endogenous (isolated cytoplasmic mRNA-transport-stimulating proteins) and exogenous (poly(A), lectins, and neoglycoproteins) factors during prenatal and postnatal rat brain and liver development, revealed marked organ-dependent differences paralleling the appearance of the poly(A)- mRNA unique in the brain.
Assuntos
Encéfalo/metabolismo , Fígado/metabolismo , Proteínas de Transporte Nucleocitoplasmático , Poli A/metabolismo , RNA Mensageiro/metabolismo , Proteínas de Ligação a RNA , Animais , Transporte Biológico , Encéfalo/crescimento & desenvolvimento , Proteínas de Transporte/metabolismo , Núcleo Celular/metabolismo , Citoplasma/metabolismo , Ativação Enzimática/efeitos dos fármacos , Glicoproteínas/farmacologia , Lectinas/farmacologia , Fígado/crescimento & desenvolvimento , Membrana Nuclear/enzimologia , Nucleosídeo-Trifosfatase , Monoéster Fosfórico Hidrolases/metabolismo , Poli A/farmacologia , Ratos , Ratos EndogâmicosRESUMO
The organization of the intranuclear elements observed in histone-depleted (2 M NaCl-extracted) HeLa cell nuclei was investigated by means of electron microscopy and two-dimensional gel electrophoresis. This work was mainly aimed at verifying whether or not an intranuclear skeleton or matrix existed, which could explain the stable attachment of RNA to the residual nuclear structure after high-salt extraction, and its three-dimensional organization. We compared the ultrastructure and the polypeptide composition of RNA-containing and RNA-depleted (RNase-treated) nuclear residues, and we visualized intermediate stages of RNase action on the intranuclear material. We showed that this material was made of two types (fibrillar and granular) of salt-resistant RNP components equally sensitive to RNase when the enzyme was used prior to high-salt extraction. At least in our material and under our experimental conditions, no intranuclear matrix could be distinguished from the residual RNP material. Our results further suggest that formation of such a matrix is a path-dependent phenomenon.
Assuntos
Núcleo Celular/ultraestrutura , Células HeLa/ultraestrutura , Ribonucleoproteínas/metabolismo , Núcleo Celular/metabolismo , DNA/metabolismo , Células HeLa/metabolismo , Humanos , Microscopia Eletrônica , Peso Molecular , RNA/metabolismo , Ribonucleases , Ribonucleoproteínas/isolamento & purificaçãoRESUMO
HIP was originally identified as a gene expression in primary liver cancers, and in normal tissues such as pancreas and small intestine. Based on gene data base homologies, the HIP protein should consist of a signal peptide linked to a single carbohydrate recognition domain. To test this hypothesis HIP and the putative carbohydrate recognition domain encoded by the last 138 C-terminal amino acids, were expressed as glutathione-S-transferase proteins (GST-HIP and GST-HIP-142, respectively). Both recombinant proteins were purified by a single affinity purification step from bacterial lysates and their ability to bind saccharides coupled to trisacryl GF 2000M were tested. Our results show that HIP and HIP-142 proteins bind to lactose, moreover the binding requires divalent cations. Thus the HIP protein is a lactose-binding lectin with the characteristics of a C-type carbohydrate recognition domain of 138 amino acids in the C-terminal region.
Assuntos
Antígenos de Neoplasias , Biomarcadores Tumorais , Expressão Gênica , Lactose/metabolismo , Lectinas Tipo C , Neoplasias Hepáticas/metabolismo , Proteínas/genética , Sequência de Bases , Metabolismo dos Carboidratos , Cátions Bivalentes , Escherichia coli/genética , Técnicas de Transferência de Genes , Glutationa Transferase/genética , Humanos , Dados de Sequência Molecular , Proteínas Associadas a Pancreatite , Proteínas/metabolismo , Proteínas Recombinantes de Fusão/metabolismoRESUMO
Previous studies have demonstrated the existence of nuclear carbohydrate binding proteins in a variety of mammalian cells with molecular masses of 35,000, 67,000, and 70,000 (CBP35, CBP67, and CBP70), which are associated with nuclear ribonucleoprotein (RNP) complexes. CBP35 consists of two domains, an amino-terminal portion that is homologous to certain regions of proteins of the heterogeneous nuclear RNP complex, and a carboxyl-terminal portion homologous to beta-galactoside-specific lectins. CBP35 it has been proposed, like the glucose-specific lectin, CBP67, to guide RNP complexes through the nuclear pore. Here we show that the exposure of mature rats to stress induces an increase in nuclear CBP35 bound to CBP67 and retained on immobilized glucose. Nuclear extracts from the livers of old rats displayed no detectable stress response. This CBP35.CBP67 association detected in rat liver is considered with respect to the CBP35.CBP70 association recently observed in HL60 cell nuclear extracts.
Assuntos
Envelhecimento/metabolismo , Antígenos de Diferenciação/metabolismo , Lectinas/metabolismo , Proteínas Nucleares/metabolismo , Ribonucleoproteínas/metabolismo , Estresse Fisiológico/metabolismo , Animais , Galectina 3 , Humanos , Masculino , Peso Molecular , Ratos , Ratos WistarRESUMO
Photodynamic therapy has emerged as an alternative to chemotherapy and radiotherapy for cancer treatment. Nanoparticles have recently been proposed as effective carriers for photosensitizers. Depending on their chemical composition, these can be used for diagnosis and therapy due to the selective accumulation of the photosensitizer in cancer cells in vitro or in tumors in vivo. Multifunctional nanoplatforms combining several applications within the same nano-object emerge as potential important theranostic tools. This review, based on the chemical nature of the nanoparticles will discuss recent advances in the area of non polymeric nanoparticles for photodynamic therapy applications.
Assuntos
Nanopartículas/uso terapêutico , Fotoquimioterapia , Fármacos Fotossensibilizantes/uso terapêutico , Animais , Ouro/uso terapêutico , Humanos , Magnetismo , Nanotubos de Carbono , Dióxido de Silício/uso terapêutico , Titânio/uso terapêutico , Óxido de Zinco/uso terapêuticoAssuntos
Núcleo Celular/ultraestrutura , Animais , Fracionamento Celular , Nucléolo Celular/ultraestrutura , Núcleo Celular/análise , Núcleo Celular/metabolismo , Cromatina/ultraestrutura , Cromossomos , DNA/análise , Replicação do DNA , Regulação da Expressão Gênica , Glicopeptídeos/análise , Células HeLa/ultraestrutura , Humanos , Microscopia Eletrônica , Transcrição GênicaRESUMO
The structure and the polypeptide composition of the nuclear shell isolated from interphase HeLa cells have been investigated and compared to those of the intranuclear material. The isolated nuclear shell contains chromatin superstructures (28-32 nm thick fibres) made of tightly packed nucleosomes that resist low ionic strength conditions and that are associated with the three nuclear lamins. Chromatin in the nuclear shell exhibits very simple chemical composition. Especially, non-histone proteins are lacking. The results presented here rule out the possibility that the nuclear shell results from contamination of lamina by intranuclear elements. They suggest that the lamins are directly involved in the specific properties and in the organization of chromatin in the nuclear shell.
Assuntos
Cromatina/análise , Histonas/análise , Membrana Nuclear/análise , Nucleoproteínas/análise , Fracionamento Celular , Núcleo Celular/análise , Cromatina/metabolismo , Cromatina/ultraestrutura , Proteínas Cromossômicas não Histona/análise , Células HeLa , Proteínas de Grupo de Alta Mobilidade/análise , Humanos , Laminas , Microscopia Eletrônica , Membrana Nuclear/metabolismo , Membrana Nuclear/ultraestrutura , Nucleossomos/ultraestrutura , Ribonucleoproteínas/análiseRESUMO
The ultrastructure and the polypeptide composition of residual nuclear substructures including nuclear matrices, nuclear ghosts, and residual envelopes were investigated by means of electron microscopy and two-dimensional gel electrophoresis. Nuclear matrices were prepared by digesting isolated nuclei with DNase I alone, followed by high-salt extraction in 2 M NaCl. Nuclear ghosts were obtained by high-salt extraction of nuclei previously digested with DNase and RNase in MgCl2-containing buffers. To prepare residual envelopes, nuclei were first digested with RNase in the presence of EDTA, then digested with Mg2+ -activated DNase I, and extracted in 2 M NaCl. The results of this comparative study support the conclusion that the intranuclear matrix is made of two distinct RNA-containing elements. One of these elements appears on ultrathin sections as a thin fibrillar network. It disappears from RNase-digested nuclei, together with numerous basic proteins, whatever the conditions of digestion. Although this element is present in extranucleolar territories, it is a major component of residual nucleoli. The second element appears as coarse-beaded fibers absent from the nucleolar areas. Its preservation in residual nuclear substructures depends on the presence of Mg2+ ions during RNase digestion of nuclei. It is enriched in two minor basic proteins of relative mass 49 000 and 70 000. The involvement of this fibrogranular element in heterogeneous nuclear RNA attachment to the nuclear matrix is discussed.
Assuntos
Núcleo Celular/ultraestrutura , Ribonucleoproteínas/análise , Fracionamento Celular , Citoesqueleto/ultraestrutura , Detergentes , Eletroforese em Gel de Poliacrilamida , Células HeLa/ultraestrutura , Humanos , Microscopia Eletrônica , Microscopia de Fluorescência , Membrana Nuclear/ultraestrutura , SaisRESUMO
CBP70 is a glycoslylated lectin that interacts through either glycan-lectin or protein-protein interactions. In addition, depending on its cellular localization, this lectin has different partners, for example, galectin-3, an 82-kDa ligand in the nucleus, or Bcl-2 in the cytoplasm. In this study, we observed the persistence of plurilocalized lectin CBP70 after two heat-shock treatments conducted either under mild conditions, i.e., incubating the cells for 1 h at 42 degrees C then for 1, 3, 5, 7, or 9 h at 37 degrees C, or harsh conditions, i.e., incubation at 42 degrees C for 1, 2, 4, 6, 8, or 10 h. By combining the information collected from biochemical, fluorocytometric, confocal, and affinity-chromatography analyses, we concluded that CBP70 persisted in HL60 cells and its N-acetylglucosamine-binding sites remained active after all the heat-shock treatments tested. These data and the previously published findings reviewed in this report concur in supporting the hypothesis that CBP70 could function as an organizer of multimeric assembly, leading to the formation of various complexes in different cellular compartments, according to the needs of the cell.
Assuntos
Temperatura Alta , Lectinas/metabolismo , Acetilglucosamina/metabolismo , Sítios de Ligação , Núcleo Celular/metabolismo , Cromatografia , Citoplasma/metabolismo , Citometria de Fluxo , Células HL-60 , Humanos , Immunoblotting , Imuno-Histoquímica , Lectinas/fisiologia , Microscopia Confocal , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Temperatura , Fatores de TempoRESUMO
Changes in DNA polymerase alpha activity accompanying tissue development have been well established in several systems. In most cases, DNA polymerase alpha activity decreases with development. Here, we report observed changes in DNA polymerase alpha activity throughout embryonic chicken brain (ECB) development. The level of DNA polymerase alpha activity was found to gradually decrease by 60% (2.3 to 0.8 nmol of [3H]dCMP incorporated/mg protein/h) between 9- and 19-day-old ECB. An enzyme-linked immunosorbent assay of DNA polymerase alpha utilizing monoclonal antibody SJK 237-71 (human KB cell DNA pol-alpha binder) also demonstrated a gradual decrease (up to 60%) of antigen over this same range of development. Analysis of DNA polymerase alpha from 11- and 19-day-old ECB by a 10 to 30% glycerol density gradient revealed a high molecular weight peak sedimenting near catalase (11.3 S) with activity at the 11th day being approximately 3-fold greater than activity at the 19th day. A Western immunoblot analysis utilizing monoclonal antibody SJK 237-71 (against human KB cell DNA polymerase alpha) showed a decrease in DNA polymerase alpha from 186 kilodaltons in 9- and 11-day ECB cell-free extracts to 120 kilodaltons in extracts from 13- to 19-day ECB. The conversion of DNA polymerase alpha from a higher to a lower molecular weight form may be a regulatory mechanism in eukaryotic DNA replication.
Assuntos
DNA Polimerase II/biossíntese , Animais , Anticorpos Monoclonais/imunologia , Encéfalo/embriologia , Encéfalo/enzimologia , Embrião de Galinha , Reações Cruzadas , DNA Polimerase II/imunologia , DNA Polimerase II/isolamento & purificação , DNA Primase , Humanos , Células KB/enzimologia , RNA Nucleotidiltransferases/isolamento & purificação , Especificidade da EspécieRESUMO
The nuclear sugar-binding components (i.e., lectin-like molecules) were analyzed using isolated and membrane-depleted nuclei after incubation in the presence of fluorescein-labeled neoglycoproteins. This analysis was performed before and during the in vitro differentiation of HL60 cells into monocytes by PMA treatment and into granulocytes by DMSO treatment. The nucleoli of undifferentiated and differentiated HL60 cells were not labeled, unlike the nucleoli of other mammalian cells studied so far. This peculiarity allowed us to quantitatively analyze by flow cytometry the changes in the lectin activity associated with the extranucleolar territories enriched in ribonucleoprotein complexes. The neoglycoprotein binding was found to be significantly lower in differentiated than in undifferentiated cells. The decrease in neoglycoprotein binding was observed within the first 24 h of DMSO or PMA treatment, just before the arrest of DNA synthesis. Taking into account that the granulocytic differentiation required 72 h of chemical treatment, the extra-nucleolar lectins might be involved in modulation of the DNA synthesis rather than in phenotypic differentiation. These data are discussed in an attempt to reconcile the association of lectins with RNP complexes and their possible involvement in modulation of HL60 cell proliferation.
Assuntos
Proteínas de Transporte/análise , Transformação Celular Neoplásica/patologia , Leucemia Promielocítica Aguda/patologia , Proteínas Nucleares/análise , Receptores de Superfície Celular , Proteínas de Transporte/metabolismo , Proteínas de Transporte/fisiologia , Diferenciação Celular/fisiologia , Núcleo Celular/química , Núcleo Celular/efeitos dos fármacos , Núcleo Celular/ultraestrutura , Transformação Celular Neoplásica/química , Transformação Celular Neoplásica/efeitos dos fármacos , DNA/análise , DNA/biossíntese , Dimetil Sulfóxido/farmacologia , Fluoresceína , Fluoresceínas , Humanos , Lectinas/análise , Lectinas/metabolismo , Lectinas/fisiologia , Leucemia Promielocítica Aguda/metabolismo , Microscopia de Fluorescência , Proteínas Nucleares/metabolismo , Proteínas Nucleares/fisiologia , Fenótipo , Acetato de Tetradecanoilforbol/farmacologia , Células Tumorais CultivadasRESUMO
Nuclear sugar-binding proteins were detected in membrane-depleted nuclei isolated from hamster BHK cells and mouse L 1210 leukemia cells by means of fluorescein-labelled neoglycoproteins. In fluorescence microscopy, the fluorescence was seen throughout the nucleus but was generally brighter over the nucleoli than over the rest of the nucleus. Flow cytofluorometry analysis demonstrated the presence of nuclear sugar-binding proteins for synthetic glycoproteins associated with different sugar residues. Among the nine neoglycoproteins used, four neoglycoproteins (namely alpha-rhamnosylated, alpha-glucosylated, N-acetyl-beta-glucosaminylated and alpha-mannosylated-6P-serum albumin) strongly labelled nuclei. Various controls strongly argue for the specificity of the nuclear labelling. The possibility that some of the sugar-binding proteins might correspond to endogenous nuclear lectins is considered.
Assuntos
Núcleo Celular/análise , Lectinas/análise , Animais , Linhagem Celular , Nucléolo Celular/análise , Nucléolo Celular/metabolismo , Núcleo Celular/metabolismo , Cricetinae , Citometria de Fluxo , Glicoproteínas/metabolismo , Rim , Leucemia L1210 , Microscopia de FluorescênciaRESUMO
This work deals with the types of nuclear skeletal structures obtained from human fibroblast nuclei isolated by different procedures. It is confirmed that, in somatic vertebrate cells, the pore complex-lamina is always observed, whereas the presence of internal nucleolar and extranucleolar residual structures depends upon the method of nuclear isolation used. Furthermore, the results reported here argue for the existence of a nucleolar skeleton different from the nucleolar matrix often observed in different cell types by other investigators. The conditions of nuclear isolation which allow us to visualize this nucleolar skeleton without any other internal residual structures are described. The attachment of the nucleolar skeleton to the lamina suggested by the present data is considered in relation to the in situ position of nucleoli near the nuclear envelope.
Assuntos
Nucléolo Celular/ultraestrutura , Núcleo Celular/ultraestrutura , Células Cultivadas , Feminino , Fibroblastos/ultraestrutura , Humanos , Microscopia Eletrônica/métodosRESUMO
The intranucleolar distribution of sugar-binding sites (i.e., lectin-like molecules) was analyzed in segregated nucleoli of actinomycin D-treated HeLa cells. The detection of sugar-binding sites was performed by incubation either of permeabilized nuclei in the presence of fluorescein-labeled neoglycoproteins or of ultrathin sections cut through in situ-fixed nuclei in the presence of gold-labeled neoglycoproteins. In the former case, the fluorescent nucleolar components were identified by comparison with the nucleolar components of similarly treated cells observed in electron microscopy. For the first time, this study reveals the presence of sugar-binding sites in both the fibrillar and the granular components of the nucleolus. In view of the data already reported on the biochemical composition of the nucleolus, some of our results led us to conclude that the nucleolar sugar-binding sites are lectin-like proteins. These proteins could be associated with preribosomes since the nucleolus is the site of both synthesis and stockage of ribosomal precursors. Some results from this study, however, show that the possibility of a relationship between some lectins and a structural component cannot be excluded.
Assuntos
Nucléolo Celular/análise , Glicoproteínas/análise , Células HeLa/análise , Sítios de Ligação , Metabolismo dos Carboidratos , Nucléolo Celular/ultraestrutura , Dactinomicina/farmacologia , Células HeLa/ultraestrutura , Humanos , Proteínas de Membrana/análise , Microscopia de Fluorescência , Proteínas de Neoplasias/análiseRESUMO
Mannose-binding sites were detected at the ultrastructural level in nuclei of lizard granulosa cells in situ by means of mannosylated ferritin. When ultrathin sections of material embedded in glycol methacrylate were incubated with mannosylated ferritin, a strong labelling was observed over nucleoli, chromatin and the external leaflet of the nuclear envelope, but no labelling was detected in the perinuclear space except for nuclear pores. This labelling was clearly inhibited when sections were incubated in a solution containing both mannosylated ferritin and a sugar-related neoglycoprotein (mannosylated serum albumin). These ultrastructural data supporting the existence of nuclear lectin-like components in reptilian cells are in agreement with our previous findings about such components in nuclei isolated from mammalian cells. Owing to the unique organization of the granular component of the nucleoli in specialized granulosa cells, we are able to show that some of the mannose-binding sites are associated with ribosomal precursors.
Assuntos
Proteínas de Transporte/metabolismo , Núcleo Celular/metabolismo , Células da Granulosa/metabolismo , Manose/metabolismo , Animais , Núcleo Celular/ultraestrutura , Feminino , Células da Granulosa/ultraestrutura , Lagartos , Lectinas de Ligação a Manose , Microscopia Eletrônica , Oócitos/metabolismo , Oócitos/ultraestruturaRESUMO
Since nuclear lectins were first characterized several years ago, six lectins have been isolated. Furthermore, the existence of nuclear glycoproteins containing N-linked complex-oligosaccharide chains or O-linked GlcNAc residues was evidenced. These latter are abundant in the nucleus and are well-studied so far. The presence of both glycoprotein and lectin in the cell nucleus led us to postulate that these two proteins could interact and play a role in some nuclear activities such as the modulation of transcription and/or nuclear cytoplasmic exchanges or by the disruption of protein-protein interactions. In such context, the recent data concerning the GlcNAc-binding activity of CBP70 argued this postulate. However, to study the possible role of a glycoprotein-lectin complex, it was critical to isolate the two partners. Because CBP70 was also a cytoplasmic protein, the lectin was isolated in both cytoplasmic and nuclear compartments in order to investigate the putative ligand in the two cellular compartments. The results obtained with cross-linking experiments on isolated and membranedepleted nuclei incubated with the CBP70 bearing an iodinatable, cleavable, photoreactive cross-linking agent (sulfosuccinimidyl 2-(p-azidosalicylamido) ethyl-1,3'-dithiopropionate) and immunoprecipitation experiments with polyclonal antibodies raised against CBP70, revealed that both nuclear and cytoplasmic CBP70 have the same 82 kDa nuclear ligand which is absent in the cytoplasmic fraction. In addition, this ligand is glycosylated, containing GlcNAc residues, and, therefore, the complex between CBP70 and the 82 kDa polypeptide could be due to a glycoprotein-lectin interaction. These results raised the possibility that nuclear glycoprotein-lectin interaction could be involved in nuclear activities.
Assuntos
Lectinas/metabolismo , Extratos Celulares , Núcleo Celular/metabolismo , Reagentes de Ligações Cruzadas , Citoplasma/metabolismo , Células HL-60 , Humanos , Ligantes , Testes de Precipitina , Ligação Proteica , Aglutininas do Germe de Trigo/metabolismoRESUMO
Some years ago, a lectin designated CBP70 that recognized glucose (Glc) but had a stronger affinity for N-acetylglucosamine (GlcNAc), was first isolated from HL60 cell nuclei. Recently, a cytoplasmic form of this lectin was described, and one 82 kDa nuclear ligand was characterized for the nuclear CBP70. In the present study, the use of Pronase digestion and the trifluoromethanesulphonic acid (TFMS) procedure strongly suggest that the nuclear and the cytoplasmic CBP70 have a same 23 kDa polypeptide backbone and, consequently, could be the same protein. In order to know the protein better and to obtain the best recombinant possible in the future, the post-translational modification of the nuclear and cytoplasmic CBP70 was analyzed in terms of glycosylation. Severals lines of evidence indicate that both forms of CBP70 are N- and O-glycosylated. Surprisingly, this glycosylation pattern differs between the two forms, as revealed by beta-elimination, hydrazinolysis, peptide-N-glycosydase F (PNGase F), and TFMS reactions. The two preparations were analyzed by affinity chromatography on immobilized lectins [Ricinus communis-l agglutinin (RCA-I), Arachis hypogaea agglutinin (PNA), Galanthus nivalis agglutinin (GNA), and wheat germ agglutinin (WGA)] and by lectin-blotting analysis Sambucus nigra agglutinin (SNA), Maackia amurensis agglutinin (MAA), Lotus tetragonolobus (Lotus), succinylated-WGA, and Psathyrella velutina agglutinin (PVA)]. Both forms of CBP70 have the following sugar moities: terminal beta Gal residues, Gal beta 1-3 GalNAc, Man alpha 1-3 Man, sialic acid alpha 2-6 linked to Gal or GalNAc; and sialic acid alpha 2-3 linked to Gal. However, only nuclear CBP70 have terminal GlcNAc and alpha-L-fucose residues. All these data are consistent with the fact that different glycosylation pattern found for each form of CBP70 might act as a complementary signal for cellular targeting.