Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UMLS:C0026764 (multiple myeloma)
 36,148 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The binding of Tnp (2,4,6-trinitrophenyl) derivatives to the Fv fragment (variable region of heavy and light chains) of the mouse myeloma IgA protein MOPC 315 was investigated by 270MHz proton nuclear magnetic resonance. Two of the haptens, Tnp-glycine and Tnp-l-aspartate, are in fast exchange with the Fv fragment, and the changes in chemical shifts for both protein and hapten resonances were determined by titrations. For the tightly binding hapten epsilon-N-Tnp-alpha-N-acetyl-l-lysine, which is in slow exchange with the Fv fragment, the changes in chemical shifts for the hapten H(3)+H(5) resonances were determined by cross-saturation. By using these data and the known structure of the combining site of protein MOPC 315 [Dwek, Wain-Hobson, Dower, Gettins, Sutton, Perkins & Givol (1977), Nature (London) 266, 31-37] the mode of binding of Tnp derivatives is deduced by ring-current calculations. The trinitrophenyl ring stacks with tryptophan-93(L) (light chain) in the ;aromatic box' formed by tryptophan-93(L), tyrosine-34(L) and phenyl-alanine-34(H) (heavy chain). Further evidence for the stacking interaction with a tryptophan residue is provided by the similarity of the optical-difference spectra observed with Tnp-aminomethylphosphonate in the presence of either the Fab fragment (light chain and N-terminal half of heavy chain) of protein MOPC 315 or tryptophan. These data show that the modes of binding of all the Tnp derivatives are very similar, despite a 100-fold range in their affinities. It is also concluded that the modes of binding of Dnp (2,4-dinitrophenyl) and Tnp derivatives to protein MOPC 315 are very similar, and that the structural basis for this is that the aromatic box is large enought to allow the trinitrophenyl ring to stack with tryptophan-93(L) while still forming hydrogen bonds to asparagine-36(L) and tyrosine-34(L).
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PMID:The binding of 2,4,6-trinitrophenyl derivatives to the mouse myeloma immunoglobulin A protein MOPC 315. 62 44

The binding of four dinitrophenyl haptens to the mouse myeloma proteins MOPC 315 IgA (immunoglobulin A) and MOPC 460IgA was studied by resonance Raman spectroscopy. Isotopic substitution with 15N and 2H was used to assign features in the resonance Raman spectra of the free haptens. Changes in each of these features on binding to the proteins could then be attributed to interactions of the proteins' binding sites with either the p-NO2 or the o-NO2/amine regions of the haptens. The interactions between a given hapten and MOPC 315 IgA are often quite distinct from those between the same hapten and MOPC 460 IgA. Moreover, for both antibodies the nature of the R side chain in a Dnp-NHR (Dnp, 2,4-dinitrophenyl) compound appears to modify the interactions between the Dnp chromophore and the protein. Thus, with the haptens studied, there is no unique set of contacts between the Dnp group and the binding site. The contacts expected between epsilon-2,4-dinitrophenyl-L-lysine and the site on MOPC 315 IgA, on the basis of a recent model for this site [Dwek, Wain-Hobson, Dower, Gettins, Sutton, Perkins & Givol (1977) Nature (London) 266, 31--37] were not detected. However, the contacts between this hapten and the site on MOPC 460 IgA were closer to those predicted by the model for MOPC 315 IgA.
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PMID:Resonance Raman-spectroscopic studies of the hapten features involved in the binding of 2,4-dinitrophenyl haptens by the mouse myeloma proteins MOPC 315 and MOPC 460. 74 21

Exposure of Fc fragments derived from human IgG1 myeloma proteins to acid pH rendered the region between the Cgamma2 and Cgamma3 domains transiently susceptible to cleavage by trypsin upon return to neutral pH. Trypsin covalently linked to Sepharose was used and two fragments derived from the Cgamma2 region and one from the Cgamma3 region were purified by column chromatography. On the basis of amino acid analysis, primary sequency data, antigenic properties, and m.w., one of the Cgamma2 fragments was shown to consist of two polypeptide chains of unequal mass joined by the inter-heavy chain disulfide bonds. The larger chain corresponded to a stretch of gamma-chain between Thr 223 and Lys 338 (Eu numbering) and the shorter chain to the section between Thr 223 and Lys 248. The other Cgamma2-fragment was a disulfide-linked dimer of the Thr 223 to Lys 338 sections of the paired gamma-chains. When this latter fragment was reduced under mild conditions it dissociated into monomers indicating that there was little or no noncovalent interactions between the Cgamma2 domains. The Cgamma3-fragment was shown to be a noncovalent dimer composed of the Glu 345 to Lys 349 sections of the two gamma-chains although some heterogeneity was apparent at the amino-terminus. Circular dichroism was used to probe the conformational relationships between the isolated domains and the parent Fc. The spectral properties of Fc could not be fully accounted for on the basis of the spectra observed for the isolated domains which suggested that inter-domain interaction might be significant in Fc.
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PMID:Structure and function of immunoglobulin domains. III. Isolation and characterization of a fragment corresponding to the Cgamma2 homology region of human immunoglobin G1. 81 64

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.
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PMID:Synthesis of immunoglobulin by substrate attached mouse myeloma cells. 81 94

The light chains, but not the heavy chains, obtained from immunoglobulin A produced by the MOPC-315 mouse myeloma bind the 2,4-dinitrophenyl (DNP) group. Specific interaction with the DNP group was determined by using several immunoadsorbents, including DNP-L-lysine-Sepharose, and elution of the adsorbed light chain by DNP-glycine. Equilibrium dialysis experiments showed that the M-315 light chain in the form of dimer (45 260 daltons) has two identical and homogeneous binding sited that bind DNP-L-lysine with an intrinsic association constant of 6.3 x 103 M-1. This is the first report, to our knowledge, in which the light chain binding data permit reliable determination of the binding constant and valency of the isolated light chain, and which suggests a predominant role for the light chain in construction of the binding site in the intact immunoglobulin molecule.
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PMID:Binding of 2,4-dinitrophenyl derivatives by the light chain dimer obtained from immunoglobulin A produced by MOPC-315 mouse myeloma. 82 Mar 73

The pKa values of the three histidine residues in the Fv fragment (variable region of the heavy and light chains) of the mouse myeloma protein MOPC 315, measured by high resolution n.m.r. (nuclear magnetic resonance), are 5.9, 6.9 and 8.2. The perturbation of the pKa of one of the histidines (pKa 6.9) on the addition of hapten and the narrow linewidth of its proton resonances suggests that it is at the edge of the combining site. References to the model of the Fv fragment [Padlan, Davies, Pecht, Givol & Wright (1976) Cold Spring Harbor Symp. Quant. Biol. 41, in the press] allows assignment of the three histidine residues, histidine-102H, histidine-97L and histidine-44L. The determination of the pKa of the phosphorus group, by 31P n.m.r., of a homologous series of Dnp- and Tnp- (di- and tri-nitrophenyl) haptens has located a positively charged residue. Molecular-model studies on the conformations of these haptens show that the residue is at the edge of the site. The model suggests that the positively charged residue is either arginine-95L or lysine-52H.
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PMID:Specificity of interactions of hapten side chains with the combining site of the myeloma protein MOPC 315. 92 46

The growth of myeloma cells in Leibovitz medium supplemented with 20% serum was limited by the depletion of glutamine. A simple modification of the Leibovitz medium by increasing the concentrations of glutamine, lysine, isoleucine, leucine, sodium pyruvate, galactose, and vitamins resulted in over 100% increase in cell growth yield. The total myeloma protein produced by the cells was increased by approximately 90% in modified Leibovitz media. Analysis of spent culture media for 19 amino acids showed that the concentrations of 8 amino acids were reduced; those of 5 amino acids were increased and the other 6 did not change significantly.
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PMID:Amino acid metabolism of myeloma cells in culture. 96 31

Fv fragments prepared from the BALB/c anti-dinitrophenyl (DNP) myeloma proteins, produced by plasmacytoma MOPC 315 and XRPC 25, were used to raise anti-idiotypic antibodies in rabbits. Anti-idiotypes to these proteins were also prepared in the syngeneic BALB/c mice by injecting the intact proteins, whereas Fv fragment failed to induce antibodies in these mice. Rabbit anti-Fv of late bleedings did not show the presence of antibodies to "framework determinants" (determinants common to all V regions of mouse immunoglobulins), and they contained precipitating anti-idiotypes whereas mouse antisera contained non-precipitating anti-idiotypes. No cross-reaction was observed between anti-idiotypes to these myeloma proteins and induced anti-DNP antibodies from BALB/c mice, when either rabbit or mouse antisera were analyzed. The anti-sera were fractionated into anti-binding site and anti-non-binding site by adsorption onto the immunogen-Sepharose column and elution by DNP lysine, followed by elution with acetic acid. Only the anti-binding sites were inhibited by hapten in their interaction with the homologous myeloma protein. The rabbit anti-Fv antibodies were predominantly (over 90%) anti-non-binding site whereas the mouse anti-idiotypes were predominantly (60 to 70%) anti-binding site.
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PMID:Preparation and subfractionation of isologous and heterologous anti-idiotypes, using FV fragments. 99 90

An analysis of red cell membrane proteins in acute and chronic lymphatic leukaemia, Hodgkin's disease, lymphosarcoma, and myeloma was carried out. The electrophoretic pattern after solubilisation in urea or SDS was examined, along with migration on cellulose acetate or acrylamide in different buffers. Protein acid, basic and neutral amino acid percentages were also determined. An increase in low molecular weight and faster anodic migration proteins was noted in the lymphoblastoses, whereas the amino acid spectrum of these proteins showed percent changes in the case of some amino acids, particularly glutamic acid, phosphoserine, lysine and histidine. The alterations observed were compared with those noted previously in other haemoblastoses, congenital haemolytic and anhaemolytic blood diseases, and endoglobular or acquired metabolic defects in a closer assessment of their significance.
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PMID:[Changes in membrane proteins in the erythrocytes of patients with hemolymphoblastosis not directly involving the erythroblastic line]. 106 86

We describe a method, called affinity partitioning, for the purification of proteins containing specific ligand binding receptor sites. This method adds specificity to the procedures for protein purification with aqueous polymer two-phase systems by introduction of a polymer derivative, coupled to an appropriate ligand. The addition of a polymer-ligand that partitions predominantly into one phase shifts the protein that binds this substance to the same phase. By performing countercurrent distribution in the presence of a polymer-ligand, the protein that binds the polymer-ligand can be separated from a heterogenous mixture. One example of affinity paritioning used dextran as the polymer-ligand. Dextran was chosen since it is a constituent of the most commonly used system for partitioning proteins. In a dextran-poly(ethylene oxide) system, concanavalin A bound dextran and partitioned predominantly into the dextran-rich phase. The addition of the specific competitor, D-mannose, displaced the partition coefficient toward unity, while the application of L-fucose, a noncompetitor, had little effect. Application of affinity partitioning to the purification of another protein required the synthesis of a specific polymer-ligand. To study this we synthesized dinitrophenyl-poly-(ethylene oxide), which binds specifically to S-23 myeloma protein. Addition of dinitrophenyl-poly(ethylene oxide) to the dextran-poly(ethylene oxide) phase system shifted the S-23 myeloma protein into the poly(ethylene oxide)-rich phase. epsilon-N-dinitrophenyl-L-lysine, by competing with binding of dinitrophenyl-poly(ethylene oxide), antagonized the latter's effect on the partition coefficient of S-23 myeloma protein. By adding various amounts of dinitrophenyl-poly-(ethylene oxide), we correlated the partition coefficient with concentration of polymer-ligand. A model of the action of polymer-ligand derivatives on the partition coefficient, derived from thermodynamic considerations, was found to be consistent with the experimental data relating the concentration of polymer-ligand and partition coefficient. Affinity partitioning should prove to be a useful complement to affinity chromatography in the purification of mixtures of proteins. Since cells and subcellular particles may be purified with aqueous polymer two-phase systems, affinity partitioning might be applied to their fractionation by using polymer-ligands specific for unique surface receptors.
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PMID:Affinity partitioning. A method for purification of proteins using specific polymer-ligands in aqueous polymer two-phase systems. 111 12


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