Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UNIPROT:P30536 (PBS)
 9,886 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

An hCG-like material has been extracted from human sperm. These experiments were designed to characterize this material. Sperms of 10 volunteers were separated from seminal fluid, washed in PBS three times, and resuspended in 0.5 ml of the same buffer. Samples were pooled; cells were disrupted by sonication and extracted in alkaline buffer by constant agitation at 4 degrees C. The extract was ultracentrifuged at 4 degrees C. Supernate was lyophilized and reconstituted in 2 cc of distilled water. This material presented a dose-response curve parallel to those of IS2-hCG and CR119 in beta hCG RIA. When chromatographed in a Sephadex G-150 column the extract eluted within the hCG range and immunoreacted in the specific beta hCG RIA. When absorbed onto a concanavalin A--Sepharose column, all recovered immunoreactive material eluted after exposure to alpha-D-methylglucoside, indicating that it is a glycoprotein. The extract stimulated progesterone and testosterone secretion in porcine granulosa cells and decapsulated rat testis, respectively, indicating its biologic potency.
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PMID:Presence of a human chorionic gonadotropin--like substance in human sperm. 57 20

Purified and desialylated glycoprotein M from human O erythrocytes precipitates with B and H specific lectin from Evonymous europaeus seeds, both in PBS and 0.2% Triton X-100. Desialylated, N-terminal fragment (MT-1) obtained by trypsin digestion of M glycoprotein does not precipitate with Evonymous lectin but inhibits precipitation.
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PMID:Localization and immunochemical characterization of the lectin Evonymous europaeus receptor site on the glycoprotein from human O erythrocytes. 74 64

T cell-deprived mice acutely infected with S. mansoni suffer microvesicular hepatocyte damage which is not seen in infected, immunological intact animals. A cationic fraction (CEF6) of the PBS-soluble portion of S. mansoni eggs (SEA) induces antibodies which, on passive transfer, prevent hepatocyte damage. CEF6 contains 2 antigens, omega 1 and alpha 1, and has also been shown to be a useful serodiagnostic reagent. This paper describes the purification and characterization of the 2 antigens present in CEF6. omega 1 is a monomeric glycoprotein with a pI greater than 9.0 and a molecular weight of 31 kDa. Alpha 1 consists of two immunologically cross-reactive dimers, 41 and 36 kDa in non-reducing conditions, each of which consists of one unique and one common glycoprotein subcomponent. In ELISA with mouse and human infection sera omega 1 is shown to be S. mansoni specific and is better able to distinguish S. mansoni infections from other schistosome infections than are unfractionated SEA, CEF6 or alpha 1. Passive transfer of monospecific anti-omega 1 sera into S. mansoni infected, T cell-deprived mice completely prevented the occurrence of microvesicular hepatocyte damage in these animals. Monospecific anti-alpha 1 serum had no hepatoprotective capacity.
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PMID:The purification, characterization, serological activity and hepatotoxic properties of two cationic glycoproteins (alpha 1 and omega 1) from Schistosoma mansoni eggs. 174 48

The identification of antibodies to platelet-specific antigens is important for correctly diagnosing neonatal alloimmune thrombocytopenia, posttransfusion purpura and refractoriness due to platelet-specific antibodies. However, the serologic identification of these platelet-specific antibodies is complicated by the presence of anti-HLA antibodies. We examined and compared the diagnostic usefulness of acid-treated and chloroquine-treated platelets for the discrimination of platelet-specific antibodies from anti-HLA antibodies. The viability of acid-treated platelets is 83.4%, which is better than that of chloroquine-treated platelets (52.6%). The antigenicity of HLA class I antigens of acid-treated platelets was significantly reduced compared with that of PBS- or chloroquine-treated platelets. On the other hand, platelet surface glycoprotein Ib and glycoprotein IIb/IIIa, and platelet-specific antigens were stable following acid or chloroquine treatment. Chloroquine-treated platelets were not suitable targets for analysis by immunofluorescence flow cytometry because of nonspecific fluorescence derived from platelet damage. We conclude that acid-treated platelets are more suitable targets than chloroquine-treated platelets for screening for platelet-specific antibodies and also for analyses of the specificity of platelet-specific antibodies.
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PMID:Acid treatment of platelets as a simple procedure for distinguishing platelet-specific antibodies from anti-HLA antibodies: comparison with chloroquine treatment. 223 61

A soluble form of the human CD4 glycoprotein (sCD4), the cellular receptor for human HIV, was treated with various physical, chemical, and enzymic regimens and tested over a range of concentrations for its capacity to inhibit the binding of HIV to CD4+ T cells. Reduction of disulfide bonds and alkylation in denaturing buffer (8 M urea) destroyed the inhibitory activity of sCD4, whereas reduction and alkylation in PBS had no effect. Derivatization or digestion of carbohydrate groups by periodate oxidation or by glycolytic enzyme digestion did not affect sCD4 inhibitory capacity. Digestion with trypsin or endoproteinase Glu-C destroyed activity. A limited digestion of sCD4 with endoproteinase Glu-C resulted in a mixture of fragments, however, and the mixture had inhibitory activity equivalent to that of intact sCD4. Within this mixture, a fragment of 23 kDa was identified that binds to HIV. Although sCD4 can be digested to yield fully active fragments, the requirement for intrachain disulfide bonding indicates that the minimum sized portion of CD4 that will retain full affinity for HIV will have to be formulated with a proper tertiary structure.
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PMID:Structural features of CD4 required for binding to HIV. 253 5

A new method was studied for eliminating HLA class I antigens from the surface of platelets without damaging the cells. Platelets were exposed to an acid solution (pH 3.0) to eliminate the antigenicity of HLA class I antigens. The reduction in antigenicities of HLA class I common antigen and individual HLA class I antigens by acid treatment was marked. Patients' sera which contained multispecific HLA antibodies reacted with PBS-treated platelets, but not with acid-treated platelets. No changes were observed in the antigenicities of glycoprotein Ib or glycoprotein IIb/IIIa. The viability of acid-treated platelets was 83%. Ultrastructural investigations revealed no significant difference between the PBS-treated platelets and acid-treated platelets. The platelet function studies showed that the aggregation of acid-treated platelets induced by various agonists was only slightly reduced compared with PBS-treated platelets. We propose that acid-treated platelets are promising for clinical use in patients refractory to platelet transfusions and may be superior to chloroquine-treated platelets for analysis of the specificity of antiplatelet antibodies.
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PMID:New approach to eliminate HLA class I antigens from platelet surface without cell damage: acid treatment at pH 3.0. 261 55

This experiment was designed (1) to determine if H-Y antigen is expressed on the cell surface of pre-implantation equine blastocyst stage embryos, (2) if so, to identify differences in expression on inner cell mass (ICM) verses trophectoderm cells and (3) to evaluate whether the detection of this glycoprotein would aid in the identification of equine embryonic sex. A total of 33 blastocyst stage horse embryos were collected 6-7 days post-ovulation by trans-cervical flush and were immediately evaluated for the presence of H-Y antigen. Additionally, 17 embryos, collected at similar stages and cultured for 72 h, were similarly evaluated. Embryos were recovered and evaluated by use of a dissecting microscope and then washed for 5 min in phosphate buffered saline supplemented with 1 g/l glucose, 36 mg/l pyruvate, 1% antibiotic-antimycotic and 10% fetal calf serum (FCS) (PBS-2). Embryos were placed in the primary antibody medium and cultured for 60 min. The primary antibody medium consisted of monoclonal antibodies to H-Y antigen (previously determined to have male-specific activity) dilute 1/5 (v/v) with PBS-2 (without FCS, PBS-1). Following an additional wash, embryos were cultured in PBS-1 containing 1/10 (v/v) fluorescein isothiocyanate conjugated goat anti-mouse or rabbit antimouse IgM Fc specific antiserum. Embryos were evaluated at 200-400 x to identify cell specific fluorescence of either trophectoderm or ICM cells. Following evaluation, embryonic sex was independently verified with karyotypes to identify sex chromosomes. Of the 50 embryos evaluated, 29 were evaluated as non-fluorescent and 21 fluorescent. Expression of H-Y antigen was detected on both trophectoderm and ICM cell types in those embryos classified as fluorescent.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Evaluation of the expression of a male-specific antigen on cells of equine blastocysts. 305 65

Searching for the best procedure for simultaneous estimation of the anterior pituitary hormones, extraction efficiencies of various media, additives such as urea and triton X-100, and physical treatments such as freezing-thawing (F-T) and sonication, were examined by measuring prolactin (PRL), growth hormone (GH), lutropin (LH), follitropin (FSH), and thyrotropin (TSH) in the extracts. Ethanolic media (60% EtOH) gave high yields of PRL at neutral to alkaline pH, but poor extraction of GH accompanied by a marked loss of its immunoreactivity during storage. Ethanolic media also gave a poor yield of LH even at high pH. Aqueous media like PBS at various pH, 0.1 M acetic acid and distilled water were considerably effective in the extraction of GH, LH, FSH and TSH if they were coupled with F-T and sonication. However, high yields of PRL could not be obtained with these aqueous media even with F-T and sonication. Hartree's 40% EtOH-6% ammonium acetate, pH 5.1, solubilized considerable amounts of glycoprotein hormones, but yielded almost no GH and only a small amount of PRL. The addition of triton X-100 to PBS (pH 7) at 0.1% resulted in the maximum extraction of glycoprotein hormones with homogenization and F-T, but further sonication was necessary for GH and PRL. When the anterior pituitaries were homogenized and frozen-thawed in PBS (pH 7) containing 1 M urea, yields of PRL, GH, LH, FSH, and TSH were maximum, and sonication did not cause any additional extraction, indicating that this procedure, i.e. homogenization and F-T in 1 M urea-PBS, would be the best for the simultaneous estimation of these anterior pituitary hormones.
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PMID:Choice of extraction procedure for estimation of anterior pituitary hormone content. 343 4

Ocular mucin, the major product of conjunctival goblet cells, constitutes the innermost layer of preocular tear film. Ocular mucin is known for its limited amount and inaccessibility. Using impression cytology, mucus strands collected from the inferior fornix of either rabbit or human eyes were found to contain inflammatory cellular debris. In order to circumvent these difficulties and to isolate native mucin molecule(s), we bathed rabbit eyes in fluid containing isotonic PBS and 5.5 X 10(-4) M acetylcholine for 4 or 12 hr. Bathing fluids containing rabbit ocular mucin (ROM), 1 ml per eye, were pooled and combined with 1M guanidine HCl and protease inhibitors containing EDTA, PMSF, and sodium azide to avoid any possible enzymatic degradation, and then separated under the same conditions by Sepharose CL-4B. In parallel, commercial porcine stomach mucin (PSM) was purified and used to compare with ROM. We also developed nitrocellulose-based dot semi-quantitative assays for nucleic acid, protein, and glycoprotein. PAS-positive fractions monitored by such a dot assay were collected at CL-4B void volume and then separated from nucleic acid contaminants by CsCl-gradient ultracentrifugation. A protein fraction, 65K, poorly-glycosylated, with high contents of Asx, Glx, and Gly was found strongly associated with both ROM and PSM, and was only separable by ultracentrifugation in 4M guanidine HCl and CsCl. Purification of the ROM was verified by SDS-polyacrylamide gel electrophoresis, amino acid analysis, and carbohydrate analysis. These results will allow future exploration of the molecular mechanism by which tear film is achieved.
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PMID:Purification and characterization of rabbit ocular mucin. 362 33

Isolated microvilli of the MAT-C1 subline of the 13762 rat mammary adenocarcinoma contain a transmembrane complex composed of a cell surface, cytoskeleton-associated glycoprotein (CAG), actin, and a 58,000-dalton polypeptide (58K). The behavior of CAG has been studied by differential centrifugation and velocity sedimentation gradient centrifugation of detergent extracts of microvilli. CAG can be pelleted along with a fraction of the microvillar actin even in the presence of ionic detergents and under microfilament-depolymerizing conditions. By velocity sedimentation analysis CAG in Triton/PBS extracts sediments as a large, heterogeneous species (sedimentation coefficient greater than 25S). In Sarkosyl and sodium dodecyl sulfate (SDS) the size and heterogeneity are somewhat reduced. In SDS CAG sediments as a 20S species in the absence of mercaptoethanol and as a 5S species in the presence of mercaptoethanol. These results indicate that CAG is a disulfide-linked multimer in the microvillus membrane. We suggest that the stable multimeric structure of CAG permits it to act as the membrane association site for several microfilaments and plays an important role in the formation and stabilization of the microvillus structure.
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PMID:Actin-associated cell-surface glycoprotein from ascites cell microvilli: a disulfide-linked multimer. 405 17


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