Amino-Acid Sequence of an Allotype b4 Light Chain from a Rabbit Antibody to Streptococcal Carbohydrate

Chen, K. C. S.; Kindt, Thomas J.; Krause, Richard M.

doi:10.1073/pnas.71.5.1995

Cited by 42 publications

(21 citation statements)

References 21 publications

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“…The profile obtained was similar to that previously reported (Cannon et al, 1978). As the rabbit k b4 light chain contains only one invariant arginine residue in the C region at position 211 (Chen et al, 1974) and one invariant arginine at position 61 in the variable region (Margolies et al, 1975), a large fragment including residues 62-211 may be isolated from this digest (see Figure 1). Additional arginine residues located amino-terminal to position 61 in heterogeneous nonimmune light chains resulted in smaller fragments that did not interfere with purification of the 62-211 peptide, which was obtained in 64% yield.…”

Section: Resultssupporting

confidence: 85%

Isolation and characterization of an active variable domain from a homogeneous rabbit antibody light chain

Ehrlich¹,

Matsueda²,

Haber³

et al. 1979

Biochemistry

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The variable domain (VL) of allotype b4 light chains of rabbit IgG was isolated from both nonimmune heterogeneous IgG and a homogeneous antibody directed against type III pneumococcal polysaccharide. Light chains were first isolated and then cleaved under mild acidic conditions between residues 109 and 110. Reduction with dithiothreitol in guanidine hydrochloride cleaved both intradomain disulfide bridges as well as the interdomain disulfide bridge joining the variable and constant domain. The sulfhydryl groups were protected after reduction by p-chloromercuribenzoate. VL was isolated from this mixture of variable and constant domains by affinity chromatography, utilizing sheep antibodies directed against a peptide including residues 110--211 from nonimmune IgG light chain. The isolated VL domain was identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and automated Edman degradation. VL from a homogeneous antibody was treated with dithiothreitol to remove p-chloromercuribenzoate, reoxidized, and recombined with homologous heavy chain. The binding of this recombinant to type III pneumococcal polysaccharide was identical with that of the light-chain--heavy-chain recombinant.

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Section: Resultssupporting

confidence: 85%

Isolation and characterization of an active variable domain from a homogeneous rabbit antibody light chain

Ehrlich¹,

Matsueda²,

Haber³

et al. 1979

Biochemistry

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“…Radiolabeled 4539 Ab was bound strongly and specifically to insolubilized anti-b4 serum (14), but did not bind to insolubilized anti-b5, anti-b6, or anti-b9 serum. This binding could be completely inhibited by unlabeled 4539 Ab or pool b4 IgG, or by 4135 Ab, which has the prototype sequence (15). These three preparations were equally effective as inhibitors over the concentration range tested.…”

Section: Resultsmentioning

confidence: 92%

“…Fig. 1 shows the CL-region sequence from position 110-145 for 4539 Ab and for the prototype b4 L chain of 4135 Ab (15). In 4539 serine is found in place of the usual alanine at position 121 and leucine in place of glutamine at position 124.…”

Section: Resultsmentioning

confidence: 99%

A genetic polymorphism in the constant region of rabbit b4 kappa chains.

Sogn¹,

Kindt²

1976

The Journal of Experimental Medicine

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Much of what is known about the number and arrangement of genes encoding immunoglobulins, and the mechanisms involved in their selective expression, derives from studies involving allotypes. Rabbit allotypes have figured prominently in these studies particularly because they are found in almost every region of the immunoglobulin molecule. Amino acid sequence studies on rabbit allotypes of group a in the Vs region and group b in the C~ region have revealed an unusual degree of structural complexity within these allotypes in that many amino acid differences are found between alternative forms (1). For example, the b4 kappa-chain sequence differs in the constant region from that of the b9 kappa chain at approximately 33% of the positions (2). A further complexity of the rabbit C~ region is that b4 light chains from homogeneous antibodies differ from one another in certain positions (3, 4).The present report describes an inherited C~-region structural variant observed among L chains with the allotype b4. This variation of allotype b4 (b4var) 1 consists of two amino acid substitutions, at residues 121 and 124. The variation has been observed in five generations of a pedigreed family. Serologic analysis has revealed that b4 TM chains are indistinguishable with regard to their b4 allotype from those with the prototype b4 sequence. Use of this variant as a genetic marker can provide information concerning the number of genes encoding light-chain constant regions. Materials and MethodsRabbit 4539 (ala3/b4b4) underwent three immunization series with Group C streptococcal vaccine using methods previously described (5). During the third series, a monoclonal response was observed and three nonsurgical exchange transfusions (6) were carried out. A homogeneous antibody with allotype a3b4 (4539 Ab) was isolated from the transfusion plasma by DEAEcellulose chromatography and chromatography on a Group C carbohydrate immunoadsorbent column (7). Amino acid sequence studies on the light chain of 4539 Ab have been carried out using a strategy described elsewhere (8). Constant-region sequence variation was first observed upon sequence determination of a peptide (residues 110-207) isolated after acid cleavage (9) of the tryptic fragment encompassing residues 62-207.Analysis of rabbit sera for the presence of this variant sequence was carried out as follows: IgG, isolated by DEAE-cellulose chromatography, was mildly reduced and alkylated. Heavy and light

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“…While these immunoglobulins have proven useful in structural (3)(4)(5) and genetic studies (2)(3)(4)(5)(6)(7)(8)(9), the immunologic mechanisms leading to their production remain unclear (2) . It is possible that large numbers of specific clones are stimulated initially, as with other antigens, but that immunologic (affinity) maturation (10,11) procedes to the extreme .…”

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confidence: 99%

Clonal dominance. I. Restricted nature of the IgM antibody response to group A streptococcal carbohydrate in mice.

Briles¹,

Davie²

1975

The Journal of Experimental Medicine

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The IgM antibody response of mice to the streptococcal group A carbohydrate (GAC) was measured. With most strains tested, large amounts of IgM antibody were produced; in AKR mice, over 1% of the total nucleated spleen cells secreted IgM anti-GAC antibody after hyperimmunization. The relative avidity of the antibody was extimated by a modification of the Jerne plaque assay where spleen cells from individual mice were tested against erythrocytes with varying GAC epitope densitymthese studies showed that the earliest, as well as latest, IgM antibodies produced were highly restricted in avidity heterogeneity. No evidence of affinity maturation was seen upon hyperimmunization. These data favor the conclusion that the restricted IgG response seen in mice hyperimmunized to GAC is not the result of affinity driven competition for antigen among precursor cells.

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Amino-Acid Sequence of an Allotype b4 Light Chain from a Rabbit Antibody to Streptococcal Carbohydrate

Cited by 42 publications

References 21 publications

Isolation and characterization of an active variable domain from a homogeneous rabbit antibody light chain

Isolation and characterization of an active variable domain from a homogeneous rabbit antibody light chain

A genetic polymorphism in the constant region of rabbit b4 kappa chains.

Clonal dominance. I. Restricted nature of the IgM antibody response to group A streptococcal carbohydrate in mice.

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