Thermodynamic and spectroscopic comparison of the binding sites of the mouse myeloma protein 315 and of its light chain dimer

FEBS Letters ◽  
1977 ◽  
Vol 78 (2) ◽  
pp. 211-215 ◽  
Author(s):  
A. Licht ◽  
D. Lancet ◽  
I. Schechter ◽  
I. Pecht
Keyword(s):  
Light Chain ◽  
Binding Sites ◽  
Myeloma Protein ◽  
Chain Dimer ◽  
Mouse Myeloma

scholarly journals Interchain disulphide-bond formation in the assembly of immunoglobulin G. Heavy-chain dimer as an intermediate

1968 ◽  
Vol 109 (4) ◽  
pp. 637-643 ◽  
Author(s):  
Brigitte A. Askonas ◽  
Alan R. Williamson
Keyword(s):  
Heavy Chain ◽  
Light Chain ◽  
Immunoglobulin G ◽  
Bond Formation ◽  
Disulphide Bond ◽  
Pulse Labelling ◽  
Myeloma Protein ◽  
Chain Dimer ◽  
Disulphide Bond Formation ◽  
Covalently Linked

1. The role of disulphide-bond formation in the assembly of G2a myeloma protein 5563 was studied by pulse-labelling ascitic plasma cells of tumour-line 5563 for 2–8min. with radioactive amino acids, and analysing the intracellular proteins. Myeloma-protein determinants were first purified by ion-exchange chromatography under conditions that do not dissociate non-covalently linked sub-units of immunoglobulin G. The pulse-labelled material was then analysed by electrophoresis on polyacrylamide gels in sodium dodecyl sulphate–phosphate–urea buffer, which dissociates non-covalently linked sub-units; after gel electrophoresis, radioactive protein bands were located by radioautography, and characterized immunologically after elution. 2. Two heavy-chain intermediates were detected: (i) heavy-chain dimer; (ii) the dimer with one light chain attached. Free light chains had previously been shown to be intermediates in assembly. No evidence for the presence of half-molecules (one light chain attached to one heavy chain) was obtained. The formation of the disulphide bond between the heavy chains thus appears to precede the light-chain–heavy-chain linkage in immunoglobulin G assembly.

scholarly journals Amino Acid Sequence of the Light Chain from a Mouse Myeloma Protein with Anti-Hapten Activity: Evidence for a Third Type of Light Chain

1971 ◽  
Vol 68 (11) ◽  
pp. 2623-2626 ◽  
Author(s):  
E. P. Schulenburg ◽  
E. S. Simms ◽  
R. G. Lynch ◽  
R. A. Bradshaw ◽  
H. N. Eisen
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequence ◽  
Light Chain ◽  
Myeloma Protein ◽  
Mouse Myeloma

scholarly journals Untranslated immunoglobulin kappa light chain mRNA in a lambda light chain-producing mouse myeloma, MOPC104E.

1980 ◽  
Vol 255 (11) ◽  
pp. 5291-5295
Author(s):  
T. Kataoka ◽  
M. Ono ◽  
M. Kawakami ◽  
Y. Ikawa ◽  
M. Aida ◽  
...  
Keyword(s):  
Light Chain ◽  
Kappa Light Chain ◽  
Lambda Light Chain ◽  
Immunoglobulin Kappa ◽  
Mouse Myeloma

scholarly journals Identification, characterization and cloning of myr 1, a mammalian myosin-I.

1993 ◽  
Vol 120 (6) ◽  
pp. 1393-1403 ◽  
Author(s):  
C Ruppert ◽  
R Kroschewski ◽  
M Bähler
Keyword(s):  
Amino Acid ◽  
Light Chain ◽  
Brush Border ◽  
Binding Sites ◽  
Neuronal Development ◽  
Amino Acid Sequences ◽  
Dependent Manner ◽  
Tail Domain ◽  
Myosin I ◽  
Splice Forms

We have identified, characterized and cloned a novel mammalian myosin-I motor-molecule, called myr 1 (myosin-I from rat). Myr 1 exists in three alternative splice forms: myr 1a, myr 1b, and myr 1c. These splice forms differ in their numbers of putative calmodulin/light chain binding sites. Myr 1a-c were selectively released by ATP, bound in a nucleotide-dependent manner to F-actin and exhibited amino acid sequences characteristic of myosin-I motor domains. In addition to the motor domain, they contained a regulatory domain with up to six putative calmodulin/light chain binding sites and a tail domain. The tail domain exhibited 47% amino acid sequence identity to the brush border myosin-I tail domain, demonstrating that myr 1 is related to the only other mammalian myosin-I motor molecule that has been characterized so far. In contrast to brush border myosin-I which is expressed in mature enterocytes, myr 1 splice forms were differentially expressed in all tested tissues. Therefore, myr 1 is the first mammalian myosin-I motor molecule with a widespread tissue distribution in neonatal and adult tissues. The myr 1a splice form was preferentially expressed in neuronal tissues. Its expression was developmentally regulated during rat forebrain ontogeny and subcellular fractionation revealed an enrichment in purified growth cone particles, data consistent with a role for myr 1a in neuronal development.

scholarly journals Biosynthesis of immunoglobulin A (IgA). Secretion and addition of carbohydrate to monomer and polymer forms of a mouse myeloma protein

1973 ◽  
Vol 136 (3) ◽  
pp. 589-596 ◽  
Author(s):  
E. Della Corte ◽  
R. M. E. Parkhouse
Keyword(s):  
Plasma Cell ◽  
Molecular Species ◽  
Specific Antibody ◽  
Early Stage ◽  
Immunoglobulin A ◽  
Carbohydrate Composition ◽  
Myeloma Protein ◽  
J Chain ◽  
Co Precipitation ◽  
Mouse Myeloma

Cell suspensions of mouse plasma-cell tumour MOPC 315 secreting predominantly IgA (immunoglobulin A) monomer and dimer were incubated with radioactive leucine, mannose, galactose and fucose for various periods of time. The amounts of secreted and intracellular immunoglobulins were measured by co-precipitation with specific antibody, and the molecular species present were assessed by electrophoresis in polyacrylamide gels. Analysis of the secreted myeloma protein demonstrated that monomer and dimer IgA molecules are identical with respect to carbohydrate composition and rate of secretion. Within the cell, the myeloma protein is almost entirely accounted for by monomer units which either leave the cell as such or are polymerized with the addition of J chain close to the time of secretion. The results support the concept of a stepwise addition of carbohydrate residues to IgA immunoglobulin during the process of secretion. Similar patterns of carbohydrate assembly were found for the monomer or dimer molecules. Mannose residues are added at an early stage, whereas fucose is added close to the time of secretion. Galactose is also added early, but some may also be incorporated at a later stage. Control of IgA polymerization is considered unlikely to reflect regulation at the level of carbohydrate addition, and it is suggested that the critical controlling factor is the J chain.

Extinction of expression of immunoglobulin genes in myeloma X fibroblast somatic cell hybrids

1987 ◽  
Vol 7 (2) ◽  
pp. 936-939
Author(s):  
A Greenberg ◽  
R Ber ◽  
Z Kra-Oz ◽  
R Laskov
Keyword(s):  
Light Chain ◽  
Immunoglobulin Genes ◽  
Somatic Cell Hybrids ◽  
Immunoglobulin Production ◽  
Myeloma Cells ◽  
Rna Transcripts ◽  
Processing Level ◽  
Early Processing ◽  
Polypeptide Chains ◽  
Mouse Myeloma

Adherent hybrids between immunoglobulin-producing mouse myeloma cells and fibroblasts do not produce immunoglobulin polypeptide chains. These hybrids retained the actively rearranged immunoglobulin genes of the myeloma parental cells but lacked immunoglobulin heavy- and light-chain RNA transcripts. We conclude that the shutoff of immunoglobulin production in these hybrids occurs at the transcription or early processing level.

scholarly journals Helper T cell recognition of the variable domains of a mouse myeloma protein (315). Effect of the major histocompatibility complex and domain conformation.

1982 ◽  
Vol 155 (6) ◽  
pp. 1587-1596 ◽  
Author(s):  
T Jørgensen ◽  
K Hannestad
Keyword(s):  
T Cells ◽  
Cell Response ◽  
Helper T Cells ◽  
Helper Cell ◽  
Spleen Cells ◽  
Myeloma Protein ◽  
Helper Cells ◽  
Low Responders ◽  
High Responsiveness ◽  
Mouse Myeloma

We have examined the recognition of the variable (V) domain of the heavy (VH) and light (V lambda 2) chains of mouse myeloma protein 315 by helper T cells. Mice were primed with the isolated V domain in complete Freund's adjuvant, and carrier (V domain)-primed spleen cells were transferred together with hapten (NIP)-primed spleen cells to recipient mice that were boosted with NIP3-Fab-315. The helper cell response to both domains was governed by H-2-linked immune response (Ir) genes, and VH-315 and V lambda 2 displayed different Ir phenotypes. H-2k conferred high responsiveness to VH on three different genetic backgrounds, BALB/c, C3H, and B10; mice of the d and b haplotypes were low responders. Conversely, H-2d conferred high responsiveness to V lambda 2 on two backgrounds, BALB/c and C3H, whereas mice of the k haplotype were low responders to this domain. Non-H-2 genes of the B10 background extinguished the helper cell response to V lambda 2 in animals with the high responder d haplotype. The VH Ir gene mapped to the K-A interval of the H-2 complex. Unfolded (completely reduced and alkylated) V domains primed helper cells as efficiently as folded domains for responses to NIP3-Fab-315, indicating that the helper cells recognized an antigenic determinant that was not conformation-dependent. The data indicate that there exists helper T cells which recognize each member of the M315 pair of V domains independent of the other, and that these V domains are recognized like conventional extrinsic protein antigens.

MONOCLONAL ANTIBODIES THAT RECOGNIZE Ca2+-INDUCED CONFORMER OF PROTEIN C, INDEPENDENT OF GLA RESIDUES

1987 ◽  
Author(s):  
T Sugo ◽  
S Tanabe ◽  
K Shinoda ◽  
M Matsuda
Keyword(s):  
Monoclonal Antibodies ◽  
Light Chain ◽  
Metal Binding ◽  
Binding Sites ◽  
Plasma Proteins ◽  
Protein C ◽  
The Other ◽  
Metal Binding Sites ◽  
Direct Elisa ◽  
Gla Domain

Monoclonal antibodies (MCA’s) were prepared against human protein C (PC) according to Köhler & Milstein, and those that recognize the Ca2+-dependent PC conformers were screened by direct ELISA in the presence of 2 mM either CaCl2 or EDTA. Out of nine MCAߣs thus screened, five MCA's designated as HPC-1˜5, respectively, were found to react with PC in the presence of Ca2+ but not EDTA. By SDS-PAGE coupled with Western Blotting performed in the presence of 2 mM CaCl2, we found that two MCA’s HPC-1 and 2, recognized the light chain, and two others, HPC-3 and 4, recognized the heavy chain of PC. But another MCA, HPC-5 was found to react with only non-reduced antigens. Further study showed that HPC-1 and 5 failed to react with the Gla-domainless PC, i.e. PC from which the N-terminal Gla-domain of the light chain had been cleaved off by α-chymotrypsin. However, all the other three MCA's retained the reactivity with the antigen in the presence of Ca2+ even after the Gla-domain had been removed. The binding of these MCA’s to PC in the presence of Ca2+ was found to be saturable with respect to the Ca2+ concentration and the half maximal binding for each MCA was calculated to be about 0.5+mM. Moreover, many other divalent cations such as Mg2+, Mn2+ , Ba2+, Zn2+, Co2+, Sr2+, were found to substitute for Ca2+ in inducing the metal ion-dependent but Gla-domain-independent conformer of PC.Cross-reactivity to other vitamin K-aependent plasma proteins was examined by direct ELISA; HPC-2 and 3 reacted solely to PC, but HPC-1 and 4 also reacted with prothrombin and HPC-5 with both prothrombin and factor X.These findings indicated that there are two or more metal binding sites besides the Gla-domain, possibly one in the light chain and the other(s) in the heavy chain. The presence of these metal binding sites may contribute to the unique conformer of vitamin K-dependent plasma proteins including protein C.

scholarly journals Crystallization of the Fv fragment of mouse myeloma protein M315

1979 ◽  
Vol 181 (2) ◽  
pp. 497-499 ◽  
Author(s):  
R Aschaffenburg ◽  
D C Phillips ◽  
D R Rose ◽  
B J Sutton ◽  
S K Dower ◽  
...  
Keyword(s):  
Ethylene Glycol ◽  
Unit Cell ◽  
Poly Ethylene Glycol ◽  
Myeloma Protein ◽  
Space Group ◽  
Cell Dimensions ◽  
Poly Ethylene ◽  
Unit Cell Dimensions ◽  
Glycol Solution ◽  
Mouse Myeloma

The Fv fragment of mouse myeloma protein M313 was crystallized from poly(ethylene glycol) solution in the form of monoclinic crystals, space group C2 and unit cell dimensions a = 5.96 nm (59.6 A), b = 5.66 nm (56.6 A), c = 13.79 nm (13.9 A) and beta = 99.7 degrees. Some unusual effects of poly(ethylene glycol)on protein crystals were noted and are discussed.

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