Restriction in IgM expression—1

Rodwell, John D.; Karush, Fred

doi:10.1016/0161-5890(80)90181-9

Cited by 26 publications

(15 citation statements)

References 23 publications

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“…For instance, H chains from high affinity antibodies, which have undergone more extensive amino acid sequence changes (due to somatic mutations), may associate less favorably than H chains from low affinity antibodies, which are more likely to have a sequence coded by germ-line genes (and resulting in associative phenomena more similar to those observed with myeloma proteins). Such a hypothesis is consistent with both the reassociation results we have seen (Table 2) and various theories concerning the role of somatic mutational events (25,26). In addition, a recent report has indicated that the third hypervariable region (especially residue 96) plays an important role in the self association of L chains (27).…”

Section: Discussionsupporting

confidence: 79%

“…The underlying principle of such a mechanism is the precommittment of a cell to the synthesis of both H and L chains that possess a high energy ofassociation and form an active site complementary to the activating antigen (i.e., the same V regions as the receptor). Postactivational mechanisms of gene regulation, such as somatic mutation (10,26), could result in the synthesis of an intact molecule that has increased binding affinity for the antigen, by using the same H and L chain subgroups as the receptor molecule. This report supports such events as the predominant or sole mechanism behind the phenomenon of affinity maturation, which was first described by Eisen and Siskind (29).…”

Section: Discussionmentioning

confidence: 99%

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Restricted reassociation of heavy and light chains from hapten-specific monoclonal antibodies.

Kranz¹,

Voss²

1981

Proc. Natl. Acad. Sci. U.S.A.

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Six murine monoclonal antifluorescyl antibody clones encompassing a defined range of affinities and containing K light chains with IgG1 or IgG2 heavy chains were examined. As the fluorescence ofthe ligand is quenched >90% when fluorescein is bound by antifluorescyl antibodies, fluorescence quenching was assayed to monitor polypeptide reconstitution and active site formation on mixing of resolved heavy (H) Two experimental approaches have generally characterized efforts to understand the organization and expression of immunoglobulin genes. One approach has involved amino acid sequence analyses of myeloma proteins (1, 2) and, more recently, hybridoma proteins (3). Another approach has been the cloning (4) and hybridization (5) of DNAs coding for these complex group ofproteins. Both approaches have contributed to current views of antibody diversity and genetics.Based on such work, antibody molecules are known to consist of two heavy (H) and two light (L) chains, each containing variable (V) and constant (C) regions. Translation of the L chain is preceded by the rearrangement of germ-line genes such that the C region is brought together with one ofmultiple V regions through a joining segment, J (6). Emerging evidence seems to indicate that a similar mechanism exists for H chains (2), although an additional region (D) located between the V and J segments also appears to contribute to diversity (7). It is believed that the number and variety of V, D, and J segment genes present in the germ line, and their combinatorial joining, generates antibody diversity and binding site specificity. Affinity of the antibody for a certain ligand could understandably, although not necessarily, be controlled by related mechanisms of gene rearrangement. Further diversity and refinement of antigen binding may be produced through somatic point mutation (8) or assortment of minigene sets or both (9).It has been shown that a given V gene can associate with any C gene controlling H chains of different classes (10). Likewise, Studies of H and L chain reassociation have involved either myeloma proteins for which the specific immunogen is unknown or a complex heterogeneous mixture ofspecific antibody molecules. The capacity of H and L chains isolated from monoclonal antibodies of the same specificity and various affinities to reassociate or form an active site has not been critically examined. The experimental approach used here has been to determine the ability of homologous and heterologous H and L chains, derived from monoclonal antifluorescyl hybridomas, not only to reassociate but also to form functional active sites. Both preferential recombination between certain H and L chains and a strict requirement for the homologous chains to form an active molecule have been observed. These results have led to a proposed mechanism that may account for the apparent restrictions involving H and L chain interactions.

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Section: Discussionsupporting

confidence: 79%

Section: Discussionmentioning

confidence: 99%

Restricted reassociation of heavy and light chains from hapten-specific monoclonal antibodies.

Kranz¹,

Voss²

1981

Proc. Natl. Acad. Sci. U.S.A.

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“…Its identity and purity were established by COOH-terminal sequence analysis with carboxypeptidase A and by analytical isoelectric focusing. Amino acid analysis of the reduced and alkylated VH confirmed that the intradomain disulfide bond was intact (13). Chickens were immunized intramuscularly with 1 mg of this fragment dissolved in saline and emulsified with complete Freund's adjuvant.…”

Section: Methodsmentioning

confidence: 99%

“…The V region fragment of the A chain of a human IgM myeloma protein was prepared as described by Rodwell and Karush (13). Its identity and purity were established by COOH-terminal sequence analysis with carboxypeptidase A and by analytical isoelectric focusing.…”

Section: Methodsmentioning

confidence: 99%

“…Although some workers have detected surface immunoglobulins on some human T cells (10)(11)(12), the relationship between these molecules and serum immunoglobulins is uncertain. In this paper we provide evidence that certain human and marmoset T cells express and synthesize a surface component that is related antigenically to the variable region of immunoglobulin heavy chain (VH) of a human Waldenstrom macroglobulin (13). This study was facilitated by the use of long-term in vitro lines of marmoset lymphocytes of strict Tor B-cell phenotype (14) that reacted with appropriate antibodies to human immunoglobulins (15).…”

mentioning

confidence: 99%

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