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1.
Incorporation of radioactive fucose into the immunoglobulin G1 myeloma protein secreted by mouse plasma-cell tumour MOPC 21 is stereospecific for the l-isomer. Heavy chains of the secreted form of the myeloma protein carry 90% of the label in fucose residues of their carbohydrate moieties. A small but significant amount of the intracellular immunoglobulin G1 of the mouse plasma-cell tumour MOPC 21 appears to be labelled. Serum in the incubation medium supplies low-molecular-weight diffusible substances necessary to maintain continuous secretion of fucose-labelled myeloma protein beyond 2-3h, and of leucine-labelled myeloma protein beyond 6-8h. In medium containing extensively dialysed serum the secretion of leucine- and fucose-labelled myeloma protein can be restored by the addition of 250mum-d-mannose, 250mum-d-galactose and 250mum-glucosamine. Synthesis and secretion appear to be facilitated in the presence of these sugars, although secretion of myeloma protein devoid of terminal fucose residues is possible for a limited time-period.  相似文献   

2.
Investigation of biosynthesis of J chain in plasmacytomas induced in NZB mice revealed that this protein was not only synthesized in the cells that produce polymer immunoglobulin A but also in those that produce immunoglobulin G monomer. It was also found that protein similar to J chain of BALB/c-mice was associated with polymer immunoglobulin A but not with immunoglobulin G of NZB mouse myeloma proteins.  相似文献   

3.
MPC 11 mouse myeloma cells synthesize two immunoglobulin kappa light chains, coded by two separate genes. One of these Kappa-chains has no variable region and is degraded intracellularly. The other is a full-length kappa-chain contaning both variable and constant regions: this chain is secreted, both by itself and combined with heavy chains in molecules of immunoglobulin G. This paper reports the amino acid sequence of the myeloma MPC 11 full-length kappa-chain. The chain is unusual in having 12 extra residues at its N-terminus when its sequence is aligned with those of other mouse kappa-chains; no other anomalies were found in its sequence.  相似文献   

4.
This study was designed to determine the time in the intracellular life of immunoglobulin when the carbohydrate moieties are added. Plasma cells from a mouse myeloma tumor were exposed to glucosamine-3H (a "bridge" sugar), galactose-3H, or leucine-3H. With each of the above isotopes, the percentage of total radioactive immunoglobulin that has been secreted after different periods of labeling and the extent to which puromycin prevented incorporation into immunoglobulin were determined. The results indicate that both galactose and glucosamine (in its N-acetyl form) become covalently incorporated into immunoglobulin G late in its intracellular life and suggest that glucosamine is also added onto nascent polypeptide chains (i.e., on polyribosomes).  相似文献   

5.
Turnover of myeloma messenger DNA   总被引:1,自引:0,他引:1  
The turnover of poly A + RNA in myeloma microsomes and postmicrosomal cytoplasm has been determined by analyzing the labeling kinetics of RNA in myeloma cells provided with a constant precursor pool of H3-uridine. The halflife of cytoplasmic poly A + RNA was found to be 12.2 hours, that of microsomal poly A + RNA 44.4 hours. Since the majority of the poly A + RNA in myeloma microsomes is immunoglobulin mRNA, the data suggest that immunoglobulin mRNA is turned over very slowly.  相似文献   

6.
Staphylococcal protein A binds efficiently to the Fc region of goat immunoglobulin G antibodies only after they are immune complexed to immobilized, but not fluid-phase, polyvalent antigen (human myeloma immunoglobulin E protein) or monovalent hapten (methotrexate). Compared to fluid-phase or immobilized free immunoglobulin G, the reactivity of anti-immunoglobulin antibodies bound to solid-phase antigen was enhanced at least 300-fold. Results with immobilized methotrexate indicated that two molecules of immunoglobulin G must be bound in proximity to bind one molecule of protein A. Thus, aggregation appears to be a necessary condition for protein A binding.  相似文献   

7.
The recombination of dimers of immunoglobulin peptide chains   总被引:13,自引:3,他引:10       下载免费PDF全文
1. Both the gamma and light peptide chains of human pooled and myeloma immunoglobulin G can be prepared as non-aggregating dimers at pH5.4 in 4mm-sodium acetate buffer. The dimeric state is maintained by non-covalent bonds, since the formation of interchain disulphide bonds was prevented by alkylation of the thiol groups. In the case of the light chains there is some evidence that the dimers are in equilibrium with a small amount of monomer. 2. When such dimers of the gamma and light chains are mixed at pH5.4 in 4mm-sodium acetate buffer they combine rapidly, yielding a product that resembles the original immunoglobulin G in its physicochemical and antigenic properties. However, the original optical rotatory dispersion spectrum was regained only with the homogeneous myeloma protein. The recombined pooled immunoglobulin G had a spectrum slightly different from the original, suggesting that at least some of the recombinant molecules had not regained native conformations. 3. Dimers of gamma chains stabilized by interchain disulphide bonds were able to recombine with light chains. However, light chains stabilized in the dimeric state by interchain disulphide bonds would not combine with gamma chains. 4. The chains of rabbit immunoglobulin G behave similarly to the human chains in this system, apart from the alkylated light chains showing clearer evidence of monomeric components.  相似文献   

8.
A model mammalian cell system for the production of recombinant proteins was investigated. Murine myeloma cells which had lost the ability to produce both heavy and light chain immunoglobulin molecules were transfected with a vector containing the immunoglobulin heavy chain promoter and enhancer elements linked to the human growth hormone gene. The growth kinetics of G32, a clonal isolate, were found to be similar to both the parent myeloma and hybridomas. However, production of hGH by G32 was growth associated, rather than as a secondary metabolite as is the case for hybridomas. In addition, G32 produced hGH at molar levels greater than most hybridomas.Abbreviations ELISA Enzyme Linked Immunosorbent Assay - Ig Immunoglobulin - MAb Monoclonal Antibody - X63 Murine Myeloma Cell Line P3X63-Ag8.653  相似文献   

9.
An improved method for the isolation of normal immunoglobulin E is described. The method is based on the use of an immunoadsorbent formed by the mechanical entrapment of antibodies against a myeloma immunoglobulin E into a lattice of a highly cross-linked macroporous polyacrylamide gel. Normal immunoglobulin E was isolated from an immunoglobulin E-rich serum pool as well as from an individual healthy donor. The isolated immunoglobulin E was contaminated with about 20% of immunoglobulin G. An antiserum against normal immunoglobulin E was prepared by immunizing rabbits with the isolated immunoglobulin E.  相似文献   

10.
Cultured mouse myeloma cells grow in suspension and synthesize and secrete large amounts of immunoglobulin. Mouse myeloma cells which attach to a plastic substratum have been obtained by mutagenesis and subsequent selection. Normal mouse myeloma cells will also attach to plastic tissue culture dishes pre-treated with poly-L-lysine. The attached cells synthesize and secrete the same large amounts of immunoglobulin as the suspended cells.  相似文献   

11.
In multiple myeloma, circulating "clonotypic" B cells, that express the immunoglobulin rearrangement of the malignant plasma cell clone, can be indirectly detected by PCR. Their role as potential "feeder" cells for the malignant plasma cell pool remains controversial. Here we established for the first time an approach that allows direct tracking of such clonotypic cells by labeling with patient-specific immunoglobulin ligands in 15 patients with myeloma. Fifty percent of patients showed evidence of clonotypic B cells in blood or bone marrow by PCR. Epitope-mimicking peptides from random libraries were selected on each patient's individual immunoglobulin and used as ligands to trace cells expressing the idiotypic immunoglobulin on their surface. We established a flow cytometry and immunofluorescence protocol to track clonotypic B cells and validated it in two independent monoclonal B cell systems. Using this method, we found clonotypic B cells in only one out of 15 myeloma patients. In view of the assay's validated sensitivity level of 10(-3), this surprising data suggests that the abundance of such cells has been vastly overestimated in the past and that they apparently represent a very rare population in myeloma. Our novel tracing approach may open perspectives to isolate and analyze clonotypic B cells and determine their role in myeloma pathobiology.  相似文献   

12.
The synthesis of immunoglobulin and of nuclear proteins has been studied in synchronized mouse myeloma cells of the C1 line. Synchronization has been obtained by a double thymidine block. C1 cells synthesize immunoglobulin at a relatively constant rate throughout the cell cycle except for mitosis, when a decrease in the rate of synthesis of total protein and of immunoglobulin is observed. Cell synchrony around mitosis is not sufficiently good to determine whether immunoglobulin is synthesized at all. Nuclear protein and in particular histones appear to be synthesized synchronously with DNA during the S phase of the cell cycle.  相似文献   

13.
IN a genetic, marker1, 2 from human γA2 immunoglobulin, it has recently been shown3 that the γA2 proteins belonging to the AM2(+) genetic variant lack the disulphide bond linking the heavy and light chains but that the other variants Am2(?) have the usual characteristics of other immunoglobulins. The fact that twenty out of twenty-two γA2 myeloma proteins (primarily from Caucasians) were Am2(+) explains previous reports that the H—L disulphide bond is absent in the γA2 molecules and that these molecules dissociate in acid. We have, however, found a third γA2 myeloma protein from a Caucasian (Rou) which failed to dissociate in the presence of acid and urea. It was typed by Dr Kunkel as an Am2(?) and the light chains are of the kappa type. The first two Am2(?) immunoglobulins identified3 were lambda type.  相似文献   

14.
Investigation of hindered rotation in carbamates reveals the high flexibility and ionic character of the CN bond as compared to common amides. This flexibility decreases in the case of thiocarbamates. The mechanism of activation of carbamates has been explored. Computations have proven the possibility of formation of an intramolecular H-bond in carbamates and thiocarbamates. This intramolecular H-bond is formed immediately after protonation of the carbamate. The possibility of formation of zwitterions as intermediates in the decomposition of carbamic and dithiocarbamic acids is discussed.  相似文献   

15.
A routine screening of monoclonal gammopathies (M.G.) was performed in the serum from 36, 015 blood donors by cellulose acetate electrophoresis. The incidence of M.G. was estimated to 0.14 per cent. About 86 per cent of cases can be classified as asymptomatic M.G. and 14 per cent as malignant M.G. (myeloma or Waldenstr?m macroglobulinemia). In asymptomatic forms, heavy chain classes are only IgG or IgM with a large predominance of IgG (86,4%). It is suggested that donors in whom M.G. have been detected should not be allowed to give blood. A yearly clinical, hematological and an immunoglobulin check-up is recommended to these patients in order to defect the first sign of a malignant process.  相似文献   

16.
An effect of salt concentration on the human myeloma immunoglobulin G structure was studied by means of circular dichroism, thermal perturbation difference spectroscopy and isoelectric focusing in a pH gradient created by a concentration gradient of glucose in borate buffer solution. Immunoglobulin G (K) Iva showed a significant shift of isoelectric point to the alkaline region as a result of the increase in salt concentration. The difference spectra indicated a change in the exposure of tyrosine residues as a result of increase in salt concentration. No changes in the circular dichroic spectra with salt concentration were observed between 205 and 250 nm. Spectral changes observed for the undigested immunoglobulin G molecule are more marked than those observed for the isolated Fab fragments.  相似文献   

17.
Free polyribosomes isolated from mouse myeloma cells in tissue culture synthesize immunoglobulin chains. The presence of these peptide chains in the cytoplasm of intact myeloma cells has been investigated. Some immunoglobulin chains were observed, but it could not be ruled out that these were originally inside cisternae of the endoplasmic reticulum, which were broken during hogenization. We have also investigated the transport of the hypothetical cytoplastic immunoglobulins into the cisternae of the endoplasmic reticulum after incubation with radioactive amino acids and subsequent chase in the absence of protein synthesis. A model to account for synthesis of immunoglobulins on free polysomes is presented. This model assigns specificity for translation on membrane-bound polysomes to the N-terminal region of secretory proteins.  相似文献   

18.
K Imaizumi  T Shiga 《Biorheology》1983,20(5):569-577
Using a rheoscope, combined with a TV image analyzer and a computer, the effects of immunoglobulins and IgG-fragments on the process of human erythrocyte aggregation were determined. The immunoglobulins accelerated the aggregation; the effect increased with their molecular weight, i.e., IgG congruent to gamma-globulin less than IgA less than IgM. An empirical relationship, expressing the dependence on the immunoglobulin concentrations, was proposed. F(ab')2-fragment accelerated the aggregation, more effectively than IgG, while Fab- and Fc-fragments did not. Therefore, two Fab-portions are presumably needed to form the aggregates, and the flexibility between two Fab-portions may be important. A case of multiple myeloma showed an increased aggregation, due to the increased myeloma protein.  相似文献   

19.
Glycoprotein-glycans have recently been implicated to play a variety of functional roles. The same glycan chain have been found complexed with proteins of diverse functions. In this article two such glycan chains found attached to Fc regions of immunoglobulin G and immunoglobulin M have been studied. An extensive simulated annealing procedure have been adopted to arrive at a low-energy minimum of the two oligosaccharides. Molecular dynamics simulations have been performed to study the flexibility of the glycosidic linkages. It was found that both glycan chains can undergo conformational transitions and adopt folded and extended conformations. The two β(1–2) linkages of complex-type glycan had been found to prefer different conformational regime and the terminal fucose linked to the GlcNAc residue drastically modifies the GlcNAc β(1–4)GlcNAc linkage conformation. In the high-mannose type glycan chain α(1–3) linkages can induce flexibility in addition to the α(1–6) linkages. The results have been compared with recent experimental nmr and fluorescence energy transfer data. © 1998 John Wiley & Sons, Inc. Biopoly 45: 177–190, 1998  相似文献   

20.
The first small-angle scattering study of an immunoglobulin A is reported. Neutron measurements have been made to determine conformational parameters of the mouse myeloma protein MOPC315 and to relate these to previous immunoglobulin G results. Use of the contrast method shows that the MOPC315 IgA molecule is not simply globular, that it has a dry volume of 220.0 +/- 4.5 nm3 corresponding to a mass density of 1.275 +/- 0.025 g cm-3 and that its full and cross-sectional radii of gyration, corrected for concentration dependence, are 7.97 +/- 0.07 nm, 2.40 +/- 0.08 nm and 1.33 +/- 0.07 nm respectively. Similar study of its Fab fragment gives a dry molecular volume of 69.0 +/- 0.7 nm3, a mass density of 1.285 +/- 0.015 g cm-3 and uncorrected radii of gyration that are consistent with those of the parent and support an overall "T" or "Y" conformation in solution. Addition to saturation of a small monovalent dinitrophenyl hapten leaves the dry volume of the whole molecule unaltered, but may slightly lower one or more of its radii of gyration. The significance of this finding is discussed. Comparative studies with rabbit anti-dinitrophenyl immunoglobulin G antibody suggest a different initial conformation but similar consequences of hapten binding, which, if real, are probably unrelated to classical complement fixation.  相似文献   

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