scholarly journals RECOMBINATION OF HEAVY AND LIGHT CHAINS OF HUMAN γA-MYELOMA PROTEINS: FORMATION OF HYBRID MOLECULES AND CONFIGURATIONAL SPECIFICITY

1966 ◽  
Vol 124 (2) ◽  
pp. 185-197 ◽  
Author(s):  
Robert A. Prendergast ◽  
Howard M. Grey ◽  
Henry G. Kunkel
Keyword(s):  
Electrophoretic Mobility ◽  
Quaternary Structure ◽  
Antigenic Specificity ◽  
Light Chains ◽  
Antigenic Analysis ◽  
Naturally Occurring ◽  
Myeloma Proteins ◽  
Hybrid Molecules ◽  
Noncovalent Bonds ◽  
Polypeptide Chains

The present studies demonstrate that the conditions necessary for reductive cleavage, isolation, and recombination of L and H polypeptide chains of human γA-myeloma globulins parallel those required for similar manipulation of the component chains of γG-globulin. Specificity of recombination was shown for chains derived from the same protein. In contrast, no intradass preferential recombination was demonstrable. Hybrid molecules, formed by reassociation of noncovalent bonds, could be synthesized from isolated chains of two immunoglobulin classes resulting in the formation of molecules of the type γA-H-γG-L and γG-H-γA-L. Several sera containing both γA- and γG-"monoclonal" peaks were studied, one of which demonstrated the L chains associated with both peaks to be identical both by electrophoretic mobility in acid-urea gel and antigenic analysis. The possibility is considered that this case represents a naturally occurring analogue of the artificially produced hybrid molecules described in this study. Configurational antigenic specificity of γA-myeloma proteins, imposed by the presence of kappa L chains in native and appropriately recombined molecules, provides a further indication of the importance of noncovalent bonds in the establishment of the quaternary structure of these proteins.

scholarly journals INDIVIDUAL ANTIGENIC SPECIFICITY OF MYELOMA PROTEINS

1965 ◽  
Vol 121 (4) ◽  
pp. 561-575 ◽  
Author(s):  
Howard M. Grey ◽  
Mart Mannik ◽  
Henry G. Kunkel
Keyword(s):  
Antigenic Specificity ◽  
Antigenic Structure ◽  
Light Chains ◽  
Antigenic Determinants ◽  
Fab Fragment ◽  
Myeloma Protein ◽  
Heavy Chains ◽  
Myeloma Proteins ◽  
The Individual ◽  
Individual Specificity

The specific antigenic structure of individual myeloma proteins was investigated for the presence of similar antigenic determinants in pooled γ-globulin and for the localization of these determinants on the γ-globulin molecules. Quantitative precipitin analyses demonstrated that in most instances absorption of antisera specific for an individual myeloma protein with large amounts of γ-globulin markedly reduced or completely removed the reactivity of the antiserum for the homologous myeloma protein. In only one instance did strong specificity remain after absorption with 100 mg of Fr II per cc of antiserum. The antigenic determinants responsible for the individual specificity were localized in all cases studied solely to the Fab fragment produced by papain digestion. After reductive cleavage, three patterns of localization were observed. Individual specificity could be localized either to; (a) isolated heavy chains, (b) isolated light chains, (c) antigenic determinants present only when light and heavy chains were recombined. After immunization with whole myeloma proteins, individual specificity was localized in part at least to the isolated heavy chain in four of six proteins studied. It was localized to the light chains in three of five type L proteins but in none of six type K proteins. In the instances where individual specificity of the myeloma protein was present on the light chains, it was shown that the Bence Jones protein from the same patient also contained the individual specificity. Immunization with isolated heavy or light chains furnished further evidence for the individual specificity of both types of chains. These studies on myeloma proteins furnished evidence concerning the portions of the γ-globulin molecule subject to individual antigenic variation. The light chains, particularly the L type and the Fd portion of the heavy chains were primarily involved. Evidence for the importance of the quaternary structure was also obtained from the necessity in some instances for light and heavy chains to be associated in order for individual specificity to be observed. The Fc fragment of the heavy chains on the other hand showed very limited variation which was related to subgroup specificity.

scholarly journals BENCE JONES PROTEINS AND LIGHT CHAINS OF IMMUNOGLOBULINS

1969 ◽  
Vol 130 (6) ◽  
pp. 1295-1311 ◽  
Author(s):  
Alan Solomon ◽  
Carla L. McLaughlin
Keyword(s):  
Light Chain ◽  
Light Chains ◽  
Bence Jones Protein ◽  
Amino Terminal ◽  
Type K ◽  
Myeloma Proteins ◽  
Immunoglobulin Molecule ◽  
Polypeptide Chains ◽  
Bence Jones Proteins

Three distinct classes of κ light polypeptide chains have been detected immunochemically by an antiserum (R185) prepared against a κ Bence Jones protein with a glutamyl amino terminal residue. This antiserum had specificity for κ light chains with glutamyl amino terminal residues and differentiated κ-chains with aspartyl amino terminal residues into two classes: the three κ-chain classes have been designated as κglu, κaspII, and κaspI. The ability of antiserum R185 to detect these antigenic differences on the intact immunoglobulin molecule, as well as on the isolated light chain or Bence Jones protein, made feasible the direct classification of type K myeloma proteins and M-macroglobulins (Waldenström). The multispecificity of the antiserum permitted the quantitation of type κglu light chains in normal, hypergammaglobulinemic, and hypogammaglobulinemic sera. Whereas the distribution of myeloma proteins and Bence Jones proteins in the κglu class correlated with the distribution of κglu chains in normal and hypergammaglobulinemic sera, the M-macroglobulins in the κglu class represented 90% of the total M-macroglobulins tested and revealed a marked divergence from the range of 24–31% of κglu immunoglobulins in normal sera. A preponderance of κglu chains was detected in the sera from patients with non-sex-linked hypogammaglobulinemia and represented 60–77% of the total type K light chain content. The controlled cleavage of a Bence Jones protein representative of each κ-chain class into its variant half and constant half made possible the localization on the light polypeptide chain, the reactive sites for which antiserum R185 had specificity. The correlations between immunochemical and structural classification of κ light chains are discussed.

Chemical Data on Pituitary Gonadotropins and Their Implication to Evolution

1982 ◽  
Vol 39 (1) ◽  
pp. 80-91 ◽  
Author(s):  
E. Burzawa-Gerard
Keyword(s):  
Amino Acid ◽  
Amino Acid Composition ◽  
Acid Composition ◽  
Quaternary Structure ◽  
Chelonia Mydas ◽  
Amino Acid Sequences ◽  
Chemical Data ◽  
Pituitary Hormone ◽  
Hybrid Molecules ◽  
Α And Β Subunits

Chemical data on gonadotropins from several vertebrate species are summarized and discussed from an evolutionary point of view. A high degree of homology has been observed between mammalian gonadotropins (LH and FSH) and thyrotropin (TSH). In non-mammalian species the existence of LH and FSH-like hormones has been demonstrated except for squamate and fish species. Especially in fish the number of GTHs is still controversial. One pituitary glycoprotein assumes various gonadotropic functions of the pituitary, and a second pituitary hormone (carbohydrate-poor) acts on fish ovarian growth. GTHs from bird, reptile, amphibian, and fish pituitaries have been purified and chemically characterized (amino acid composition, carbohydrate content). The existence of a quaternary structure has been demonstrated for several tetrapod LHs and fish GTHs. The amino acid composition of α and β subunits purified from turkey (Meleagris gallopavo), and turtle (Chelydra serpentira, Chelonia mydas) LHs and from common carp (Cyprinus carpio) and sturgeon (Acipenser stellatus) GTHs showed homology with the mammalian α and β subunits. The partial sequences of carp GTH subunits have shown that the carp GTH β was more closely related to mammalian LH β than to FSH β. Hybrid molecules could be obtained by association of heterologous subunits. The kinetics of subunit association has been studied in vitro. As compared to ovine LH, subunit association of carp GTH was more rapid and thermodependent. The subunit β seemed to determine the thermodependence. The various GTH subunits in living vertebrate probably derive from a common ancestral molecule.Key words: vertebrate gonadotropins, chemical characterizations, GTHs subunits, amino acid sequences, hybrid molecules, evolution.

Naturally Occurring Human Antibodies with Specificity for Light Chains of Immunoglobulins

Vox Sanguinis ◽  
1977 ◽  
Vol 32 (2) ◽  
pp. 69-76
Author(s):  
Rudolf Scherz ◽  
Rolf Pflugshaupt ◽  
René Bütler
Keyword(s):  
Light Chains ◽  
Human Antibodies ◽  
Naturally Occurring

scholarly journals Functional roles of two polypeptide chains that compose an antigen-specific suppressor T cell factor.

1984 ◽  
Vol 159 (4) ◽  
pp. 1096-1104 ◽  
Author(s):  
M Taniguchi ◽  
T Tokuhisa ◽  
T Itoh ◽  
M Kanno
Keyword(s):  
T Cell ◽  
Light Chain ◽  
Mrna Translation ◽  
Functional Expression ◽  
Light Chains ◽  
Antigen Specificity ◽  
Suppressor Factor ◽  
Functional Roles ◽  
T Cell Factor ◽  
Polypeptide Chains

The functional roles of the two polypeptide chains that compose the T cell suppressor factor (TsF) that mediates the antigen-specific and genetically restricted suppressor function were studied by using the heavy or light chains isolated from the conventional TsF or the 11S and 13S mRNA translation products of TsF. Either the heavy or the light chain of mRNA translation products reconstitutes the active TsF that suppresses the antibody response in an antigen-specific and genetically restricted manner when it is combined with the isolated heavy or light chain from the conventional TsF. As a consequence, the antigen-binding heavy chain mediates the antigen specificity of TsF. On the other hand, the I-J-positive light chain works as an element to determine the genetic restriction specificity. Thus, the identity of the histocompatibility between the I-J haplotypes on the light chain and the responding cell is essential for the functional expression of TsF. No genetic preference, however, was observed, in the association of the heavy and light chains of TsF.

Kinetic Studies on Antibody–Hapten Reactions. II. Reactions with Antibodies and their Polypeptide Chains

1973 ◽  
Vol 51 (10) ◽  
pp. 1355-1364 ◽  
Author(s):  
K. A. Kelly ◽  
A. H. Sehon ◽  
A. Froese
Keyword(s):  
Thin Layer ◽  
Rate Constant ◽  
Rate Constants ◽  
Kinetic Studies ◽  
Light Chains ◽  
Gel Chromatography ◽  
Equilibrium Studies ◽  
Polypeptide Chains

Kinetic and equilibrium studies were performed on the reactions of the hapten ε-dinitrophenyl-lysine with specific intact antibodies, reduced, alkylated, and polyalanylated antibodies, and reduced, alkylated, and polyalanylated γ-chains. No reaction was detected between the hapten and light chains. The γ-chains were found to have 0.5 combining sites per chain, and thin layer gel chromatography revealed that they existed as monomers. The rate constant of association for the reaction of γ-chains with hapten was found to be almost 1000 times lower than that for the corresponding reaction with the parent antibody. Differences in the rate constants of dissociation were much less pronounced. These results suggested that the combining site in the separated γ-chain had undergone a change in conformation.

scholarly journals Proteolytic inactivation of human factor VIII procoagulant protein by activated human protein C and its analogy with factor V

Blood ◽  
1984 ◽  
Vol 63 (2) ◽  
pp. 486-489 ◽  
Author(s):  
CA Fulcher ◽  
JE Gardiner ◽  
JH Griffin ◽  
TS Zimmerman
Keyword(s):  
Factor Viii ◽  
Protein C ◽  
Human Factor ◽  
Activated Protein C ◽  
Light Chains ◽  
Factor V ◽  
Human Factor Viii ◽  
Thrombin Activation ◽  
A Site ◽  
Polypeptide Chains

Abstract Purified human factor VIII procoagulant protein (VIII:C) was treated with purified human activated protein C (APC) and the loss of VIII:C activity correlated with proteolysis of the VIII:C polypeptides. APC proteolyzed all VIII:C polypeptides with mol wt = 92,000 or greater, but not the doublet at mol wt = 79–80,000. These results and our previous thrombin activation studies of purified VIII:C, are analogous with similar studies of factor V and form the basis for the following hypothesis: activated VIII:C consists of heavy and light chain polypeptides [mol wt = 92,000 and mol wt = 79–80,000 (or 71–72,000), respectively] which are similar in Mr to the heavy and light chains of activated factor V. Thrombin activates VIII:C and V by generating these polypeptide chains from larger precursors and APC inactivates both molecules by cleavage at a site located in the heavy chain region of activated VIII:C and V.

scholarly journals THE EMERGENCE OF ANTIBODIES WITH EITHER IDENTICAL OR UNRELATED INDIVIDUAL ANTIGENIC SPECIFICITY DURING REPEATED IMMUNIZATIONS WITH STREPTOCOCCAL VACCINES

1970 ◽  
Vol 131 (6) ◽  
pp. 1169-1189 ◽  
Author(s):  
Klaus Eichmann ◽  
Dietmar G. Braun ◽  
Ten Feizi ◽  
Richard M. Krause
Keyword(s):  
Electrophoretic Mobility ◽  
Specific Antibody ◽  
Carbohydrate Antigen ◽  
Unrelated Individual ◽  
Antigenic Specificity ◽  
Preparative Electrophoresis ◽  
Detectable Antibody ◽  
Electrophoretic Mobilities ◽  
Distinct Individual

Electrophoretically monodisperse antibody components in rabbit antisera to the carbohydrates of the Groups A and C streptococci have been examined for their individual antigenic specificity. In these antibody components which were isolated by preparative electrophoresis, individual antigenic specificity was confined to the specific antibody and was absent in the nonantibody γ-globulin. Radioprecipitation experiments and the use of immune absorbent columns constructed from goat anti-antisera, which had been absorbed with fraction II, revealed that all the specific antibody in an electrophoretically monodisperse component was reactive with the homologous anti-antibody. Antibodies with either identical or distinct individual antigenic specificities may occur in the same rabbit with repeated immunizations. Antibodies with identical antigenic specificity had identical electrophoretic mobility, whereas antibodies with unrelated antigenic specificities had distinct electrophoretic mobilities. In the interval between immunizations, if antibody to the carbohydrate antigen was absent, there was no detectable antibody with individual antigenic specificity.

Inhibition of Fibrin Monomer Polymerization by Lambda Myeloma Globulins

Blood ◽  
1972 ◽  
Vol 39 (2) ◽  
pp. 210-223 ◽  
Author(s):  
Morton Coleman ◽  
Edward M. Vigliano ◽  
Marc E. Weksler ◽  
Ralph L. Nachman
Keyword(s):  
Anticoagulant Activity ◽  
Enzymatic Digestion ◽  
Thrombin Time ◽  
Clot Retraction ◽  
Fibrin Polymerization ◽  
Myeloma Proteins ◽  
Fibrin Monomer ◽  
Low Concentrations ◽  
Polypeptide Chains

Abstract Blood obtained from seven patients with lambda type myeloma proteins showed evidence of gelatinous bulky clots, impaired clot retraction, and circulating anticoagulant activity associated with interference of fibrin monomer polymerization. Five patients had γG1 and two had γA1 myeloma proteins. The pathologic plasmas and isolated myeloma proteins had anticoagulant activity that prolonged both thrombin and reptilase times, that was not absorbed by BaSO4 or neutralized by protamine sulfate, and that resisted heating to 56°C for 10 min. Addition of excess calcium partially corrected the anticoagulant effect. The anticoagulant activity of the isolated whole myeloma proteins, the enzymatic fragments, and polypeptide chains was measured by a thrombin time assay and a spectophotometric system with fibrin monomer. Low concentrations of the isolated IgGL proteins inhibited fibrin polymerization. The IgAL proteins did not demonstrate this activity at low concentrations but were active at concentrations comparable to in vivo levels. F(ab')2 and Fab fragments produced from the IgG proteins by enzymatic digestion possessed full inhibitory activity of the native intact proteins. Fc fragments and isolated polypeptide chains did not display significant anticoagulant activity. The results suggest that the Fab sites of certain lambda myeloma proteins may bind to fibrin during clotting and fibrin polymerization.

scholarly journals Isolation and characterization of phytoferritin from pea (Pisum sativum) and Lentil (Lens esculenta)

1978 ◽  
Vol 171 (2) ◽  
pp. 349-356 ◽  
Author(s):  
R R Crichton ◽  
Y Ponce-Ortiz ◽  
M H J Koch ◽  
R Parfait ◽  
H B Stuhrmann
Keyword(s):  
Amino Acid ◽  
Pisum Sativum ◽  
Quaternary Structure ◽  
Polyacrylamide Gel Electrophoresis ◽  
Lens Esculenta ◽  
Molecular Weights ◽  
Isolation And Characterization ◽  
Angle Scattering ◽  
Scattering Study ◽  
Polypeptide Chains

Ferritin was isolated from the seeds of pea (Pisum sativum) and lentil (Lens esculenta). The homogeneity of the phytoferritins was established by polyacrylamide-gel electrophoresis. The subunit molecular weights were respectively 20 300 and 21 400 for hte pea and lentil proteins. A neutron low-angle scattering study established the molecular weight of the oligomer as 480 000 for pea apoferritin and 510 000 for lentil apoferritin. Although the quaternary structure of 24 polypeptide chains is preserved, the phytoferritins have a larger cavity in the interior than mammalian ferritins and can thus potentially store 1.2-1.4 times as much iron. The amino acid composition of the phytoferritins show some similarities to those of mammalian apoferritins; tryptic ‘fingerprinting’ reveals that there are many differences in the amino acid sequence of plant and mammalian apoferritins.

Sign in / Sign up

Export Citation Format

Share Document