Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: UNIPROT:O95477 (membrane-bound)
 29,236 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

We describe the bulk purification of a water-soluble form of RT1-A class I MHC antigens from aqueous extracts of DA liver. Using a combination of monoclonal antibody affinity, lentil lectin affinity, and gel permeation chromatography, we were able to obtain large quantities of pure water-soluble RT1-A antigens. Typically, from 40 DA livers, 0.5 mg of pure antigen with antigen activity equivalent to 7 X 10(9) nucleated DA spleen cells was obtained. The water-soluble RT1-A molecule had a discrete heavy chain of 40 kD, linked noncovalently to beta 2 microglobulin. The heavy chain of the water-soluble RT1-A molecule was 5 kD smaller than the membrane-bound form of RT1-A from DA liver membranes. The smaller molecule probably represents a secreted form of RT1-A class I molecules that lack the transmembrane domain (exon 5). Large quantities of this water-soluble RT1-A class I antigen from the DA strain, given intravenously to PVG recipients of DA cardiac allografts by a variety of protocols, did not have any effect on graft survival. The fairly ready availability of milligram quantities of pure class I transplantation antigens should be of considerable value for studies in transplantation.
 ...
PMID:Bulk purification of a naturally occurring soluble form of RT1-A class I major histocompatibility complex antigens from DA rat liver, and studies of specific immunosuppression. 329 12

The synthesis of membrane-bound and secreted immunoglobulin was investigated in the human line LICR-LON-HMy2, a cell line often used for the derivation of human hybridomas. PAGE-SDS analysis of immunoprecipitates obtained from 35S-methionine labelled cell lysates shows that LICR-LON synthesize a hitherto undetected membrane form of IgG (with a heavy chain of mol. wt 62,000) in addition to the secretor form of IgG already described (55,000 heavy chain). Tunicamycin treatment, pulse-chase experiments and Western blot analysis showed that both chains are synthesized as independent proteins. Hybridomas obtained after fusion of LICR-LON and human peripheral blood lymphocytes retained the ability of the parental cell line to synthesize gamma m and gamma s. Some of these hybrids synthesize and secrete IgM which presumably originates from the parental B-lymphocytes. Precipitation and PAGE-SDS analysis of membrane proteins after iodination of intact cells revealed only one heavy chain band, corresponding in size to that of the gamma m. No indication of the synthesis of the membrane form of IgM was found in the hybrids. These data show that the parental (lymphoid) phenotype (m and s-IgG) is codominant with the more differentiated phenotype (s-IgM) of the fusion partner cell (plasma cell). These observations are compatible with a class-specific m-s regulation operating on a different chromatin structure at the expressed Ig loci of each parental chromosome.
 ...
PMID:The regulation of membrane-bound and secreted immunoglobulins in the human lymphoid cell line LICR-LON and human hybridomas. 365 99

The RT1.A (H-2K,D type) class I major histocompatibility complex (MHC) antigens of the rat are well recognized as membrane-bound glycoproteins. In this report, we demonstrate that liver and kidney in the DA rat strain contain large amounts of a water-soluble RT1.A class I molecule with a discrete heavy chain approximately 5 kd smaller than the membrane-bound form. An identical molecule could be identified in DA rat serum. This small class I molecule carries all of the polymorphic antigenic determinants of the RT1.A av1 class I molecule. The water-soluble molecule is readily denatured in its pure form when frozen and thawed, but this does not occur when it is mixed with serum, presumably because of a stabilizing interaction with one or more carrier proteins. The half-life of the class I molecule in serum was measured to be approximately 1.5 h. The LEW rat strain produced detectable but substantially smaller amounts of water-soluble RT1.A molecules. Our studies indicate that RT1.A class I MHC antigens are synthesized and presumably secreted in a smaller water-soluble form by liver, kidney, and possibly other tissues under physiological conditions, a point of considerable interest in view of the immunoregulatory functions of the membrane-bound forms of these molecules.
 ...
PMID:Water-soluble form of RT1.A class I MHC molecules in the kidney and liver of the rat. 381 9

Chicken lymphocyte membrane immunoglobulins (Ig), were precipitated with mouse monoclonal antibodies specific for heavy and light chain isotypes and analyzed by polyacrylamide gel electrophoresis. Very little or no membrane-bound IgG and IgA was detected. After sequential precipitation and removal of IgM reactive with any of three monoclonal anti-mu antibodies, anti-light chain antibody precipitated residual Ig with a relative electrophoretic mobility similar to that of IgM. Under reducing conditions, these surface Ig molecules had a heavy chain that appeared slightly larger (approximately 81,000 daltons) than mu-chain (approximately 79,000 daltons), and light chains of approximately 25,000 daltons. Complete clearance of membrane-bound IgM reactive with an anti-mu allotype antiserum left similar molecules precipitate by monoclonal anti-light chain antibody. These non-IgM molecules could be detected on the surface of lymphocytes from blood, spleen, bursa and the B cell line RAV-1, but not from thymus or blood from an agammaglobulinemic chicken. After capping of B cell surface IgM with anti-mu, immunofluorescent staining with anti-light chain antibody revealed residual Ig molecules disturbed across the surface of more than 90% of the IgM-bearing cells. The data suggest the existence of an avian homologue of mammalian IgD. Affinity-purified goat anti-mu antibodies and a fourth monoclonal anti-mu antibody reacted with both IgM and the putative IgD molecules, which suggests that the IgD homologue shares at least one common determinant with chicken IgM.
 ...
PMID:Evidence for an IgD homologue on chicken lymphocytes. 618 38

The heavy chain genes for IgM (C mu) and IgD (C delta) are expressed differentially during B cell maturation and activation. We have determined the role that transcription plays in the regulation of these changes by using the method of in vitro nascent RNA chain elongation. In neonatal cells that express much lower densities of IgD than IgM on their surface, transcription of C delta is observed at half the level of C mu. This 3:1 transcriptional ratio of mu to delta is preserved in mature resting cells, which express higher densities of IgD on the surface than IgM. When activated by the mitogen, lipopolysaccharide (LPS), transcription of C mu is preferentially enhanced. However, C delta transcription is not shut off even though the expression of IgD in the stimulated cells is greatly decreased. In all three differentiative stages, polymerase unloading occurs in the vicinity of a large inverted repeat sequence, 5' to C delta and 3' to the mu membrane exons. This suggests that the developmental selection of secreted vs. membrane-bound carboxyl-terminal exons is controlled by RNA cleavage. The data presented here, together with our previous analysis of mRNA and protein synthesis, show that the differential expression of IgM and IgD in normal B lymphocytes is regulated at the transcriptional, translational, and posttranslation levels.
 ...
PMID:Transcriptional regulation of the mu-delta heavy chain locus in normal murine B lymphocytes. 620 82

The B cell lymphoma I.29 consists of a mixture of cells expressing membrane-bound immunoglobulin M (IgM) (lambda) and IgA (lambda) of identical idiotypes. Whereas most of the cells express either IgM or IgA alone, 1 to 5% of the cells in this tumor express IgM and IgA simultaneously within the cytoplasm and on the cell membrane (R. Sitia et al., J. Immunol. 127:1388-1394, 1981; R. Sitia, unpublished data). When IgM+ cells are purified from the lymphoma and passaged in mice or cultured, a portion of the cells convert to IgA+. These properties suggest that some cells of the I.29 lymphoma may undergo immunoglobulin heavy chain switching, although it is also possible that the mixed population was derived by a prior switching event in a clone of cells. We performed Southern blotting experiments on genomic DNAs isolated from populations of I.29 cells containing variable proportions of IgM+ and IgA+ cells and on a number of cell lines derived from the lymphoma. The results were consistent with the deletion model for heavy chain switching, as the IgM+ cells contained rearranged mu genes and alpha genes in the germ line configuration on both the expressed and nonexpressed heavy chain chromosomes, whereas the IgA+ cells had deleted both mu genes and contained one rearranged and one germ line alpha gene. In addition, segments of DNA located within the intervening sequence 5' to the mu gene, near the site of switch recombination, were deleted from both the expressed and the nonexpressed chromosomes. Although mu genes were deleted from both chromosomes in the IgA+ cells, the sites of DNA recombination differed on the two chromosomes. On the expressed chromosome, Smu sequences were recombined with S alpha sequences, whereas on the nonexpressed chromosome, Smu sequences were recombined with S gamma 3 sequences.
 ...
PMID:Rearrangements and deletions of immunoglobulin heavy chain genes in the double-producing B cell lymphoma I.29. 629 Aug 69

The murine tumor line 70Z/3 resembles a pre-B lymphocyte in containing the heavy chain of IgM (mu) as a cytoplasmic protein in the apparent absence of light chain (L). However, these cells can be induced by lipopolysaccharide to differentiate into a B lymphocyte-like state, containing mu2L2 tetramers as membrane-bound molecules. This is a accompanied by an increase in mu synthesis, the acquisition of complex carbohydrate by mu, and the induction of L chain. We wished to determine which of these events is critical for membrane deposition of mu. We found that uninduced 70Z/3 cells, as well as lipopolysaccharide-uninducible variants, contained a low, constitutive level of membrane bound mu, all of which was found as mu2L2. Dextran sulfate, another inducing agent, apparently caused a redistribution of this pre-existing surface mu without altering the pattern of mu synthesis or processing. One lipopolysaccharide-uninducible variant showed a small subset of surface mu-positive cells, and the proportion of these cells increased with a prolonged induction period. The increase in mu synthesis was nearly normal, but mu did not acquire complex carbohydrate. However, the delayed appearance of surface mu-positive cells was paralleled by a delayed increase in L chain, which occurred only in those cells with mu on their membrane. We concluded that L chain signals the transport of mu to the cell surface.
 ...
PMID:The requirement of light chain for the surface deposition of the heavy chain of immunoglobulin M. 640 41

We have used a genetic approach to study the differentiation of B lymphocytes. The cultured murine cell line 70Z/3 resembles pre-B cells in containing the heavy chain of the immunoglobulin IgM, mu, as an internal protein in the absence of light chain, L. However, overnight incubation with the B cell mitogen lipopolysaccharide (LPS) induces the cells to mature to a B lymphocyte-like state by the induction of L chain synthesis and the appearance of IgM on the cell surface. We have used immunoselection against surface-bound IgM to isolate LPS uninducible variants of 70Z/3. These fall into two complementation groups, LPS A and LPS B. LPS A variants predominated and were found at a frequency of 1/1200. These cells were completely unresponsive to LPS. LPS B was represented by a single variant in which a subset of cells was induced to display wild-type levels of membrane-bound IgM, and the proportion of induced cells increased with prolonged incubation with LPS. We detected no structural defects in either variant group, but LPS B may represent a defect in the decision to differentiate in response to LPS.
 ...
PMID:LPS-nonresponsive variants of mouse B cell lymphoma, 70Z/3: isolation and characterization. 641 57

Membrane-bound polysomes were prepared from the posterior silk gland of the silkworm, Bombyx mori, on the fourth to fifth day in the fifth larval instar. The polysomes, when supplemented with a soluble fraction from the posterior silk gland, exhibited the elongation reaction of the growing polypeptide-chains, but the initiation reaction of polypeptide synthesis was not demonstrated in this system. The predominant products synthesized on the membrane-bound polysomes were fibroin heavy chain (H-chain) and light chain (L-chain), while polypeptides of heterogeneous size classes were synthesized on the 105,000 X g-sedimentable polysomes. A substantial fraction of the fibroin L-chain synthesized was bound to the H-chain by disulfide bond. Most of the newly synthesized fibroin H- and L-chains on the membrane-bound polysomes were proved to be present within microsomal membrane vesicles because of their insensitivity to digestion with proteases in the absence of Triton X-100.
 ...
PMID:Predominant synthesis of fibroin heavy and light chains on the membrane-bound polysomes prepared from the posterior silk gland of the silkworm, Bombyx mori. 652 Jan 17

Antigen-binding receptor (ABR) molecules have been selectively radiolabeled and isolated from immunized chicken spleen cells. The specific radiolabeling of the receptors has been accomplished by utilizing a novel technique employing lactoperoxidase (LPO) covalently linked to antigen (Ag) for which human gammaglobulin was used. The cell surface ABRs were first bound to the Ag-LPO conjugates through specific recognition sites on the Ag portion of the conjugates. The bound LPO portions were then allowed to catalyze the radioiodination of the ABRs. After radiolabeling, cells were solubilized with detergents. ABRs still bound to Ag-LPO conjugates were directly isolated from the lysates via immunoaffinity chromatography utilizing an immunoaffinity reagent directed toward the antigen portion of the ABR-Ag-LPO complex. The radioactive materials isolated in this way were then analyzed via SDS-PAGE under reducing conditions. It appears that as expected, most of the specifically-labeled and isolated materials were immunoglobulin (Ig). Both the membrane-bound form of the heavy chain (mol. wt 77 K) as well as the secreted form (mol. wt 67 K) were detected, along with the light chain (mol. wt 25 K). An additional polypeptide of mol. wt 55 K was also selectively labeled and isolated along with the Ig. This may be a molecule closely associated with the membrane immunoglobulin on B-cell surface.
 ...
PMID:A novel method for radiolabeling antigen-binding receptors of lymphocytes. 660 58


<< Previous 1 2 3 4 5 6 7 8 9 10 Next >>