A new variable region in mouse immunoglobulin lambda light chains.

Sanchez, Pierre; Cazenave, Pierre‐André

doi:10.1084/jem.166.1.265

Cited by 27 publications

(8 citation statements)

References 18 publications

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“…Interestingly, 13F6-1-2 contains a rare Vλ x light chain which is observed in only 0.5% of antibody sequences and is barely detectable in normal mouse serum [17][18][19] . Vλ x , first described in 1987 [17][18][19] , is conserved among mice, humans, and other mammals 20 , and displays at least three features unique from Vλ1 and Vλ2.…”

Section: Introductionmentioning

confidence: 99%

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Complex of a Protective Antibody with Its Ebola Virus GP Peptide Epitope: Unusual Features of a Vλx Light Chain

Lee

Kuehne

Abelson

et al. 2008

Journal of Molecular Biology

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Abstract13F6-1-2 is a murine monoclonal antibody that recognizes the heavily glycosylated mucin-like domain of the Ebola virus virion-attached glycoprotein (GP) and protects animals against lethal viral challenge. Here we present the crystal structure, at 2.0 Å, of 13F6-1-2 in complex with its Ebola virus GP peptide epitope. The GP peptide binds in an extended conformation, anchored primarily by interactions to the heavy chain. Two GP residues, Gln P406 and Arg P409, make extensive side chain hydrogen bond and electrostatic interactions to the antibody and are likely critical for recognition and affinity. The 13F6-1-2 antibody utilizes a rare Vλ x light chain. The three light chain complementarity determining regions (CDRs) do not adopt canonical conformations and represent new classes of structures distinct from Vκ and other Vλ light chains. In addition, although Vλ x had been thought to confer specificity, all light chain contacts are mediated through germline-encoded residues. This structure of an antibody that protects against the Ebola virus now provides a framework for humanization and development of a post-exposure immunotherapeutic.

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Section: Introductionmentioning

confidence: 99%

“…Vλ x , first described in 1987 [17][18][19] , is conserved among mice, humans, and other mammals 20 , and displays at least three features unique from Vλ1 and Vλ2. The amino acid sequence is only ~30-33% homologous to other known Vλ and/or Vκ light chains.…”

Section: Introductionmentioning

confidence: 99%

Complex of a Protective Antibody with Its Ebola Virus GP Peptide Epitope: Unusual Features of a Vλx Light Chain

Lee

Kuehne

Abelson

et al. 2008

Journal of Molecular Biology

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“…2 to 5) (18,28). It must be emphasized that three of the probes cross-hybridize with two of the A genes because of sequence homologies; V2 reacts with VA2 and VA1, and C2 reacts with CX2 and CA3.…”

Section: Vzmentioning

confidence: 99%

“…Interestingly, the lambda genes rearrange in restricted combinations, VA1 only with CX1 or CX3 and VX2 almost exclusively with CX2 but very rarely with CA1 or CX3 (7). The newly discovered VXx gene has so far been seen rearranged only with CX2 (6,28). Recently, the order and orientation of A genes, except for Vx, were determined by deletion mapping with unique probes (18).…”

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Physical linkage of mouse lambda genes by pulsed-field gel electrophoresis suggests that the rearrangement process favors proximate target sequences.

Storb¹,

Haasch²,

Arp³

et al. 1989

Mol. Cell. Biol.

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The first complete map of a mammalian immunoglobulin gene locus is presented. Mouse A genes were mapped by pulsed-field gel electrophoresis. The gene order is V2-Vx-C2-C4-Vl-C3-Cl. The distance between V2 or Vx and the C2-C4 cluster is 74 or 55 kilobases (kb), respectively, whereas that between Vl and C3-C1 is only 19 kb; V2 and C3-Cl are at least 190 kb apart. Thus, the distances between the A subloci are inversely proportional to their frequencies of rearrangement. The related gene A5 is not within the 500 kb of the A locus mapped here.Mouse A genes have been found in four separate clusters of DNA clones which so far were physically unlinked: a cluster of clones containing the JC3 and JC1 genes, one of JC2 and the pseudogene JC4, and separate groups of clones for Vl and V2 (2, 31). Interestingly, the lambda genes rearrange in restricted combinations, VA1 only with CX1 or CX3 and VX2 almost exclusively with CX2 but very rarely with CA1 or CX3 (7). The newly discovered VXx gene has so far been seen rearranged only with CX2 (6,28). Recently, the order and orientation of A genes, except for Vx, were determined by deletion mapping with unique probes (18). The genes were all found to be in the same transcriptional orientation, and the order was determined to be V2-C2-C4-Vl-C3-Cl (18). This organization explains why V1-C2 rearrangements were not found but gives no clues about why rearrangements of Vl are about 5 to 10 times more frequent than rearrangements of V2. Furthermore, the location of Vx in this scheme was unknown. Since the sequence of Vx is as related to VK as to VX1-VX2 (6), its evolutionary association with the A locus presumably occurred in a way different from that of V1-V2 and thus its location and orientation may be unusual.We have cloned, in phage and cosmids, approximately 140 kilobases (kb) of DNA making up the VA and CA genes and their flanking regions. However, it was impossible to link up the A genes by chromosomal walking because of the paucity of unique sequences in this locus (18). We therefore used pulsed-field gel electrophoresis (PFGE) (3,30,33) transferred into lysis buffer (0.5 M EDTA, pH 9.5, 1% N-lauryl sarcosine, 0.25 mg of proteinase K per ml). The agarose inserts were incubated for 48 h at 50°C, washed extensively for 48 h in 10 mM Tris (pH 7.2)-i mM EDTA (TE) at 4°C, and stored at 4°C in TE. DNA inserts were digested in the restriction enzyme buffer specified by the manufacturer (New England BioLabs, Inc.) containing 4 mM spermidine. Each 40-,lI insert was incubated in 40 ,ul of 2x reaction mixture with 60 U of enzyme at 37°C overnight; on the next morning, a further 40 U of enzyme, 4 ,umol of spermidine, and sufficient lOx buffer and bovine serum albumin to bring these reagents to lx were added. Incubation was continued for 4 to 6 h at 37°C, and the inserts were rinsed with 3 ml of electrophoresis buffer (0.25x TAE [16]) for 30 min on ice and electrophoresed as indicated in the figure legends.5-Azacytidine treatment. 5-Azacytidine treatment was for 32 h in culture (9, 12); cel...

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cDNA clones encoding immunoglobulin λ chains from rabbit expressing the phenotype c7

et al. 1990

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A cDNA library derived from spleen cells of an unimmunized rabbit expressing the c7 phenotype of Ig lambda chains (c7+, c21-) was screened with V lambda or C lambda probes of a lambda light chain bearing c21 epitopes. The nucleotide sequences of three hybridizing clones were found to be identical within the V lambda, J lambda and C lambda regions. The V lambda region was 97% similar to that of the functional germ-line gene V lambda 2, and the C lambda region was identical to that of gene C lambda 6, recently identified. Gene C lambda 6 exhibited four codon differences when compared with gene C lambda 5, the latter encoding c21 epitopes. The data presented here and in the accompanying report (Jaton, J.-C. et al., Eur. J. Immunol. 1990, 20:2713) support the view that gene C lambda 6 encodes the C region of c7 lambda chains and that c7 and c21 markers designate two distinct isotypic forms of lambda chains. On the basis of comparative Southern blotting analyses and restriction maps of cloned genomic regions containing V lambda and C lambda genes, a scheme is proposed to account for the c7- and c21- phenotypes.

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A new variable region in mouse immunoglobulin lambda light chains.

Cited by 27 publications

References 18 publications

Complex of a Protective Antibody with Its Ebola Virus GP Peptide Epitope: Unusual Features of a Vλx Light Chain

Complex of a Protective Antibody with Its Ebola Virus GP Peptide Epitope: Unusual Features of a Vλx Light Chain

Physical linkage of mouse lambda genes by pulsed-field gel electrophoresis suggests that the rearrangement process favors proximate target sequences.

cDNA clones encoding immunoglobulin λ chains from rabbit expressing the phenotype c7

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