Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.5.1.52 (PNGase F)
 1,527 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

A new murine IgA mAb (JKT.M1), developed against Jurkat T cells chronically infected with HIV IIIB induces in vitro homotypic aggregation in several hemopoietic cell lines. The JKT.M1 Ag is expressed on a wide variety of cell types including human lymphocytes, monocytes, platelets, RBC, human umbilical vein endothelial cells, many T cell lines, myelomonocytic cell lines, and a primate kidney cell line. The JKT.M1 Ag shows differential expression on myelomonocytic cells; it is present on K562 and HL60 cell lines, which represent precursors of E and monocytes, respectively, but is not expressed on the surface of U937 and THP-1 cell lines, which appear to represent intermediate cell types of the monocytic cell lineage. However, the JKT.M1 Ag is expressed on mature peripheral blood monocytes and the MonoMac cell line. Immunoprecipitation from cell lysates (Jurkat, SupT1, PBMC, MonoMac) with the JKT.M1 mAb yields a 20-kDa Ag with few if any carbohydrate residues as determined by N-glycanase and neuraminidase treatments. The pI appears acidic by two-dimensional gel analysis, and the nonreduced form migrates more slowly than the reduced form when analyzed by SDS-PAGE suggesting the presence of intramolecular disulfide bridge(s). JKT.M1 mAb-induced cell adhesion is shown to be divalent cation- and temperature-dependent. The adhesion induced by JKT.M1 mAb is inhibited by 20 microM cytochalasin B and also by 2 mM 2-deoxyglucose plus 10 mM sodium azide suggesting that cytoskeletal changes and metabolic energy are required. Aggregation induced by JKT.M1 appears to be independent of CD43, CD44, and VLA4 (CD29/CD49d), mAb against which have also been shown to induce homotypic cell adhesion. Anti-CD18 mAb have been shown to inhibit homotypic aggregation in other studies but failed to do so in the present study. Thus JKT.M1-induced adhesion also appears to be independent of CD18, the beta-chain of leukocyte integrins. However, like mAb against LFA-1, immobilized JKT.M1 stimulates a T cell line to undergo dramatic morphologic changes which could be enhanced by the addition of phorbol ester. These data suggest that the novel 20-kDa molecule recognized by the JKT.M1 mAb may trigger cell adhesion through a previously undescribed mechanism.
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PMID:A monoclonal antibody against a novel 20-kDa protein induces cell adhesion and cytoskeleton-dependent morphologic changes. 138 18

Culture medium conditioned by phorbol 12-myristate 13-acetate-differentiated THP-1 cells contained interleukin 1 (IL-1) antagonist activity as measured by inhibition of both IL-1 beta binding to receptors on YT cells and inhibition of IL-1/phytohemagglutinin-stimulated IL-2 synthesis by LBRM-33-1A5 T cells. Based on their ability to compete for 125I-IL-1 beta binding to receptors on YT cells, four distinct antagonist proteins were purified from THP-1 cell conditioned medium using a combination of ion-exchange, hydrophobic interaction, and size exclusion chromatographies. The four proteins had different isoelectric points with molecular masses in the range 22-26 kDa and had similar specific activities for inhibition of IL-1 beta binding to cell surface receptors (Ki values 0.33-0.64 nM) and for inhibition of IL-1/phytohemagglutinin-stimulated IL-2 synthesis by 1A5 cells (IC50 values 25-100 pM). Amino-terminal sequence analysis of the two major forms (25 kDa/pI 5.1 and 22 kDa/pI 5.8) revealed complete identity for the first 27 residues in both forms. Based on the results of peptide mapping, amino acid compositional analysis and immune blotting, all of the forms were deduced to be variants of a common protein. Deglycosylation of the antagonist proteins with N-glycanase converted them to a common form (22 kDa/pI 5.8), indicating that the four isoforms represent glycosylation variants of a common protein and that asparagine-linked oligosaccharides are responsible for the observed size and charge heterogeneity.
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PMID:Purification and characterization of interleukin 1 receptor level antagonist proteins from THP-1 cells. 214 61

We describe the isolation of human fibronectin receptors (integrins) from two nonadherent promonocytic cell lines and from peripheral blood monocytes. Integrins purified from U-937 and THP-1 cells exhibited identical electrophoretic migrations on sodium dodecyl sulfate gels run under reducing (approximately Mr 150,000) and nonreducing (alpha, Mr 160,000; beta, Mr 130,000) conditions. Treatment of U-937 or THP-1 cells with phorbol esters induced these cells to express different integrins with electrophoretic mobilities (alpha, Mr 140,000; beta, Mr 115,000, nonreduced) identical to those from normal human peripheral blood monocytes. Receptors isolated from uninduced, nonadherent promonocytic leukemia cells (U-937 and THP-1) were distinct from glycoproteins IIb and IIIa and from leukocyte adhesion molecules (p150/95). However, receptors isolated here did react with an antibody known to block cell adhesion to fibronectin. The differences observed in apparent molecular masses of fibronectin receptors from uninduced and induced U-937 or THP-1 cells are removed by treatment of purified integrins with endoglycosidase F or N-glycanase. In summary, the data presented here demonstrate the purification of integrins by fibronectin affinity chromatography from human leukemia cells and normal peripheral blood monocytes. Our results suggest that these receptors differ in immature and mature monocytic cells, and are altered by glycosylation in the course of cellular maturation.
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PMID:Alteration of fibronectin receptors (integrins) in phorbol ester-treated human promonocytic leukemia cells. 297 31

The human promyelocytic leukemia ell line, HL-60, synthesized a class of high-molecular-weight (M.W. 5000 to 7000), N-linked glycopeptides as the major class of protein-bound carbohydrates. Small glycopeptides (M.W. 2500 to 3500), typical of most mammalian cells except erythrocytes, represented a minor component in these cells. The large glycopeptides were labeled efficiently with fucose, glucosamine, and galactose but only poorly with mannose. They were found not to be glycolipids, glycosaminoglycans, or mucin-type glycopeptides and were not susceptible to exoglycosidases, but they were partially degraded by endo-beta-galactosidases. These characteristics are similar to those of the large glycopeptides synthesized by erythrocytes, by another human myeloid leukemia cell line (K562), and by human and murine teratocarcinoma cells. High-molecular-weight glycopeptides predominated on another human myeloid leukemia cell line KG1, but they were expressed at low levels on both a human monocytic leukemia cel line (THP-1) and a human T-lymphoblastoid cell line (Jurkat). When HL-60 cells were induced to differentiate into macrophage-like cells with phorbol esters, the proportion of large glycopeptides decreased, and the production of small glycopeptides predominated. This shift was observed within the first several hr after exposure to phorbol esters and was temporally related to the acquisition of adherent properties by the induced cells. In contrast, when HL-60 cells were induced to differentiate into granulocytes by dimethyl sulfoxide, hypoxanthine, or retinoic acid, they continued to synthesize glycopeptides similar to uninduced cells. Human peripheral blood granulocytes synthesized primarily large glycopeptides, whereas monocytes and lymphocytes synthesized mostly small glycopeptides. These results indicate that the synthesis of high-molecular-weight glycopeptides is a property of human myeloid leukemia cell lines and that it persists throughout myeloid differentiation. A proportionate decrease in the synthesis of these large glycopeptidase is a part of the differentiation program for monocytes and macrophages.
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PMID:Decreased synthesis of high-molecular-weight glycopeptides in human promyelocytic leukemic cells (HL-60) during phorbol ester-induced macrophage differentiation. 694 6

We previously reported that the immunohistochemically defined LFA-1 antigen (LFA-1-like antigen) was expressed on various exocrine tissues uninvolved with tumors in patients with malignant diseases using LFA-1 alpha-specific monoclonal antibodies (mAb) 2F12 and HVS6B6. In this study we investigated differences at the molecular level between LFA-1 on leukocytes and LFA-1-like antigen on MKN45.16, a subline derived from an adenocarcinoma line MKN45 that expresses a high amount of LFA-1-like antigen. LFA-1-like antigen was reactive to mAb 2F12 or HVS6B6, but not to any of the other five different LFA-1 alpha (CD11a)-specific or four LFA-1 beta (CD18)-specific mAb. mAb 2F12 immunoprecipitated a 200-kDa membrane protein (LFA-1-like antigen) from MKN45.16 cells, whereas it precipitated 180-kDa (LFA-1 alpha) and 95-kDa (LFA-1 beta) proteins from a monocytic cell line (THP-1) under both nonreducing and reducing conditions. The molecular difference was confirmed further by N-glycanase treatment of the immunoprecipitates. The isoelectric point of LFA-1-like antigen was 6.0, whereas those of LFA-1 alpha and LFA-1 beta were 6.0 and 4.7, respectively, by two-dimensional gel electrophoresis. Expression of LFA-1 alpha gene on MKN45.16 cells was not detected at the mRNA level by six different sets of LFA-1 alpha-specific oligonucleotides and reverse transcription-polymerase chain reaction. These results indicated that LFA-1-like antigen on an adenocarcinoma cell line was a distinct molecule from LFA-1 on leukocytes.
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PMID:Immunohistochemically defined lymphocyte function-associated antigen 1 (LFA-1) on an adenocarcinoma cell line is a distinct molecule from LFA-1 on leukocytes. 749 27

It has been documented that human monocytes/macrophages are reactive with antibodies directed to carcinoembryonic antigen (CEA) and non-specific cross-reacting antigens (NCAs), a group of glycoproteins antigenically cross-reactive with CEA, yet the molecules responsible for this antigenic activity have not been fully clarified. In the present study, among 7 myelomonocytic cell lines tested, 2 monoblastoid lines, U-937 and THP-1, were found to express NCA-50/90, a glycosylphosphatidylinositol-anchored cell-adhesion molecule chiefly expressed on granulocytes. The 2 cell lines showed a reaction pattern with 5 distinct anti-CEA and anti-NCA monoclonal antibodies, similar to that of CHO transfectants expressing recombinant NCA-50/90. Immunoprecipitation and SDS-PAGE analyses identified glycoproteins of about 95 and 55 kDa in U-937 and THP-1 cells, respectively. Deglycosylation of the 2 antigens with N-glycanase gave the same apparent molecular mass of about 45,000, which was also the same as that of the deglycosylated form of the recombinant NCA-50/90. Upon Northern-blot analysis, only one band of approximately 2.5 kb was detected in both cell lines with a cDNA probe for NCA-50/90, which has a broad specificity to the CEA gene family members. cDNA cloning demonstrated that the 2.5-kb clones encode the peptide of NCA-50/90. The expression of NCA-50/90 by U-937 and THP-1 was down-regulated at both the protein and mRNA levels during cell differentiation from monoblastoid to monocyte/macrophage-like cells induced by stimulation with phorbol 12-myristate 13-acetate. Our observations suggest that NCA-50/90 is a differentiation antigen of cells of the monocyte/macrophage lineage as well as of the granulocyte lineage.
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PMID:Characterization of a species of non-specific cross-reacting antigen (NCA) expressed by human monocytic cell lines: structure and expression during cell differentiation. 811 77

A marked degree of macrophage infiltration of white adipose tissue (WAT) occurs in obesity and may link excess adiposity with the chronic inflammatory state underlying metabolic syndrome and other comorbidities of obesity. Excess deposition of fat in the intra-abdominal vs. subcutaneous WAT depots is a key component of metabolic syndrome. Through construction and differential screening of a murine ob/ob WAT cDNA library, we identified Slc37a2, a novel sugar transporter of the major facilitator superfamily, to be twofold enriched in intra-abdominal vs. subcutaneous fat. We find Slc37a2 is a macrophage-enriched transcript. In murine tissues, Slc37a2 transcript is restricted to spleen, thymus, and obese WAT. It is also readily detected in the RAW264.7 macrophage cell line and increases 46-fold during macrophage differentiation of THP-1 human monocytes. Compared with wild-type mice, Slc37a2 transcript is increased epididymal ninefold in ob/ob WAT and assessment of expression of the macrophage marker emr1 indicated upregulation of Slc37a2 transcript in macrophages populating ob/ob WAT. Studies with PNGase F and tunicamycin reveal the Slc37a2 protein is posttranslationally modified by addition of N-linked glycans. Slc37a2 protein migrates as heterogeneous species of approximately 50-75 kDa and its ectopic expression in mammalian cells results in the appearance of large intracellular vacuoles. We postulate that the function of this macrophage-specific putative sugar transporter is central to the metabolism of the macrophage population specifically present in obese WAT.
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PMID:The major facilitator superfamily member Slc37a2 is a novel macrophage- specific gene selectively expressed in obese white adipose tissue. 1735 11

Staphylococcal superantigen-like proteins (SSL) show no superantigenic activity but have recently been considered to act as immune suppressors. It was previously reported that SSL5 bound to P-selectin glycoprotein ligand-1 (PSGL-1) and matrix metalloproteinase (MMP)-9, leading to inhibition of leukocyte adhesion and invasion. These interactions were suggested to depend on sialic acid-containing glycans of MMP-9, but the roles of sialic acids in the interaction between SSL5 and MMP-9 are still controversial. In the present study, we prepared recombinant glutathione S-transferase-tagged SSL5 (GST-SSL5) and analyzed its binding capacity to MMP-9 by pull-down assay after various modifications of its carbohydrate moieties. We observed that GST-SSL5 specifically bound to MMP-9 from a human monocytic leukemia cell line (THP-1 cells) and inhibited its enzymatic activity in a concentration-dependent manner. After MMP-9 was treated with neuraminidase, its binding activity towards GST-SSL5 was markedly decreased. Furthermore, recombinant MMP-9 produced by sialic acid-deficient Lec2 mutant cells showed much lower affinity for SSL5 than that produced by wild-type CHO-K1 cells. Treatment of MMP-9 with PNGase F to remove N-glycan resulted in no significant change in the GST-SSL5/MMP-9 interaction. In contrast, the binding of GST-SSL5 to MMP-9 secreted from THP-1 cells cultured in the presence of an inhibitor for the biosynthesis of O-glycan (benzyl-GalNAc) was weaker than the binding of GST-SSL5 to MMP-9 secreted from untreated cells. These results strongly suggest the importance of the sialic acid-containing O-glycans of MMP-9 for the interaction of MMP-9 with GST-SSL5.
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PMID:Role of sialic acid-containing glycans of matrix metalloproteinase-9 (MMP-9) in the interaction between MMP-9 and staphylococcal superantigen-like protein 5. 2932 25