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Erratum to “Gangliosides, Ab1 and Ab2 antibodies. II. Light versus heavy chain: An idiotype–anti-idiotype case study” [Mol. Immunol. 44 (2007) 1015–1028]
Molecular Immunology 47 (2010) 939–940
Contents lists available at ScienceDirect
Molecular Immunology journal homepage: www.elsevier.com/locate/molimm
Erratum
Erratum to “Gangliosides, Ab1 and Ab2 antibodies. II. Light versus heavy chain: An idiotype–anti-idiotype case study” [Mol. Immunol. 44 (2007) 1015–1028] Alejandro López-Requena a,∗ , Yoan Machado b , Ariel Talavera c , Yuniel Fernández-Marrero a a
Department of Antibody Engineering, Center of Molecular Immunology, P.O. Box 16040, Havana 11600, Cuba Analytical Unit, Center of Molecular Immunology, P.O. Box 16040, Havana 11600, Cuba c Department of Computational and Structural Biology, Center of Molecular Immunology, P.O. Box 16040, Havana 11600, Cuba b
a r t i c l e
i n f o
Article history: Received 17 August 2009 Accepted 28 August 2009 Available online 14 November 2009
The authors regret that in the above published paper an erroneous sequence was reported, corresponding to the variable domains of a monoclonal antibody (mAb). Due to what evidently was a mishandling of samples during the cloning or sequencing steps, the sequences of the light chain variable domain (VL) of two anti-idiotypic antibodies (4F2 and 4C8 mAbs) were claimed to be identical. Now, while checking by mass spectrometry (MALDITOF-TOF, data not shown) the integrity of the purified 4F2 mAb, the tryptic peptides corresponding to the light chain were not related to those expected from the reported VL sequence. We also found a difference in the N-terminal peptide of the heavy chain variable domain (VH). As previously reported, we confirmed now by mass spectrometry that 4F2 mAb shares with anti-idiotypic 1E10 mAb (Vázquez et al., 1998) the same heavy chain (LópezRequena et al., 2007b), except for this newly found mutation that according to the mass calculations corresponded to a single residue change. These two antibodies, together with 4C8 mAb, were generated (Vázquez et al., 1998) against P3 mAb (Vázquez et al., 1995), which is specific for N-glycolyl-containing gangliosides. The previous erroneous result led us then to the conclusion that in the case of 1E10 mAb, heavy chain rules binding to P3 mAb, as VLs of 1E10 and 4F2 mAbs appeared to be quite different while heavy chains were identical (López-Requena et al., 2007b). The GenBank entry number for 4F2 VL, AY787870, has been updated with the correct sequence, obtained from a new preparation of RNA from the hybridoma cells producing 4F2 mAb and partially confirmed by mass spectrometry. Although we currently have some new, unpublished evidences from mutagenesis studies indicating that the above mentioned conclusion is not absolutely wrong, we still
DOI of original article:10.1016/j.molimm.2006.03.004. ∗ Corresponding author. Fax: +53 7 2720644. E-mail address: [email protected] (A. López-Requena). 0161-5890/$ – see front matter © 2010 Elsevier Ltd. All rights reserved. doi:10.1016/j.molimm.2009.08.031
think that the right 4F2 mAb VL sequence sets an interesting scenario. The only differences between 1E10 and 4F2 mAbs reside in two amino acid residues, one at the heavy chain framework 1 (HFR1) (also confirmed by sequencing) and the other at the light chain CDR3 (L-CDR3); specifically, both the alanine 9 residue of VH and the glycine 91 residue of VL (Kabat numbering) in 1E10 (Pérez et al., 2001) are replaced by valine residues in 4F2, which additionally constitute mutations with respect to their most closely related VH and VL germ-line gene sequences reported in the Immunogenetics Database (IMGT, http://imgt.cines.fr). The change at 4F2 L-CDR3 seems to be enough for a number of differences between these antibodies: (i) a higher affinity of 4F2 for P3 mAb (Vázquez et al., 1998); (ii) no effect on the reactivity with 4F2 mAb of the two single and the double arginine-by-serine mutations at the heavy chain CDR3 (H-CDR3) of P3 mAb (López-Requena et al., 2007b), while the double mutation did affect in some extent the binding of 1E10 mAb (López-Requena et al., 2007a); (iii) the finding of 1E10-specific peptidic phagotopes that did not bind 4F2 mAb (Pérez et al., 2002; López-Requena et al., 2007b), suggesting that they were recognizing the 1E10 VL. In contrast, and also according to the model of the P3/1E10 (idiotype/anti-idiotype) complex obtained using the crystal structure of the chimeric P3 antibody Fab fragment (Talavera et al., Mol. Immunol., 2009), it is highly improbable that the amino acid replacement at H-FR1 could provide a satisfactory explanation for the differential binding properties of 1E10 and 4F2 mAbs. In fact, according to the model of the P3/1E10 complex, the L-CDR3 glycine 91 residue is tightly packed at the VH/VL interface; thus, the model predicts that a valine residue at that position would induce a conformational rearrangement of L-CDR3 in order to fit this bulkier amino acid. This rearrangement could lead to a better shape complementarity and a larger interacting surface between P3 and 4F2 mAbs, with respect to P3 and 1E10. Therefore, these data indicate that a single amino acid difference at L-CDR3 can have important implications for the antibody reactivity, even though other results suggest
940
A. López-Requena et al. / Molecular Immunology 47 (2010) 939–940
that in the binding to its antigen (in this case, another antibody) the heavy chain seems to have a preponderant role. We are currently attempting to crystallize the P3/1E10 complex to determine the definitive topology of this idiotype/anti-idiotype interaction. References López-Requena, A., Mateo de Acosta, C., Moreno, E., González, M., Puchades, Y., Talavera, A., Vispo, N.S., Vázquez, A.M., Pérez, R., 2007a. Gangliosides, Ab1 and Ab2 antibodies I. Towards a molecular dissection of an idiotype–anti-idiotype system. Mol. Immunol. 44, 423–433. López-Requena, A., Rodríguez, M., Mateo de Acosta, C., Moreno, E., Puchades, Y., González, M., Talavera, A., Valle, A., Hernández, T., Vázquez, A.M., Pérez, R., 2007b. Gangliosides, Ab1 and Ab2 antibodies II. Light versus heavy chain: an idiotype–anti-idiotype case study. Mol. Immunol. 44, 1015–1028. Pérez, A., Lombardero, J., Mateo, C., Mustelier, G., Alfonso, M., Vázquez, A.M., Pérez, R., 2001. Immunogenetic analysis of variable regions encoding Ab1 and ␥-type
Ab2 antibodies from the NeuGc-containing ganglioside family. Hybridoma 20, 211–221. Pérez, A., Mier, E.S., Vispo, N.S., Vázquez, A.M., Pérez, R., 2002. A monoclonal antibody against NeuGc-containing gangliosides contains a regulatory idiotope involved in the interaction with B and T cells. Mol. Immunol. 39, 103–112. Talavera, A., Eriksson, A., Okvist, M., López-Requena, A., Fernández-Marrero, Y., Pérez, R., Moreno, E., Krengel, U., 2009. Crystal structure of an anti-ganglioside antibody, and modelling of the functional mimicry of its NeuGc-GM3 antigen by an anti-idiotypic antibody. Mol. Immunol., doi:10.1016/j.molimm.2009.07.032. Vázquez, A.M., Alfonso, M., Lanne, B., Karlsson, K.-A., Carr, A., Barroso, O., Fernández, L.E., Rengifo, E., Lanio, M.E., Álvarez, C., Zeuthen, J., Pérez, R., 1995. Generation of a murine monoclonal antibody specific for N-glycolylneuraminic acid-containing gangliosides that also recognizes sulfated glycolipids. Hybridoma 14, 551– 556. Vázquez, A.M., Pérez, A., Hernández, A.M., Macías, A., Alfonso, M., Bombino, G., Pérez, R., 1998. Syngeneic anti-idiotypic monoclonal antibodies to an anti-NeuGccontaining ganglioside monoclonal antibody. Hybridoma 17, 527–534.
Contents lists available at ScienceDirect
Molecular Immunology journal homepage: www.elsevier.com/locate/molimm
Erratum
Erratum to “Gangliosides, Ab1 and Ab2 antibodies. II. Light versus heavy chain: An idiotype–anti-idiotype case study” [Mol. Immunol. 44 (2007) 1015–1028] Alejandro López-Requena a,∗ , Yoan Machado b , Ariel Talavera c , Yuniel Fernández-Marrero a a
Department of Antibody Engineering, Center of Molecular Immunology, P.O. Box 16040, Havana 11600, Cuba Analytical Unit, Center of Molecular Immunology, P.O. Box 16040, Havana 11600, Cuba c Department of Computational and Structural Biology, Center of Molecular Immunology, P.O. Box 16040, Havana 11600, Cuba b
a r t i c l e
i n f o
Article history: Received 17 August 2009 Accepted 28 August 2009 Available online 14 November 2009
The authors regret that in the above published paper an erroneous sequence was reported, corresponding to the variable domains of a monoclonal antibody (mAb). Due to what evidently was a mishandling of samples during the cloning or sequencing steps, the sequences of the light chain variable domain (VL) of two anti-idiotypic antibodies (4F2 and 4C8 mAbs) were claimed to be identical. Now, while checking by mass spectrometry (MALDITOF-TOF, data not shown) the integrity of the purified 4F2 mAb, the tryptic peptides corresponding to the light chain were not related to those expected from the reported VL sequence. We also found a difference in the N-terminal peptide of the heavy chain variable domain (VH). As previously reported, we confirmed now by mass spectrometry that 4F2 mAb shares with anti-idiotypic 1E10 mAb (Vázquez et al., 1998) the same heavy chain (LópezRequena et al., 2007b), except for this newly found mutation that according to the mass calculations corresponded to a single residue change. These two antibodies, together with 4C8 mAb, were generated (Vázquez et al., 1998) against P3 mAb (Vázquez et al., 1995), which is specific for N-glycolyl-containing gangliosides. The previous erroneous result led us then to the conclusion that in the case of 1E10 mAb, heavy chain rules binding to P3 mAb, as VLs of 1E10 and 4F2 mAbs appeared to be quite different while heavy chains were identical (López-Requena et al., 2007b). The GenBank entry number for 4F2 VL, AY787870, has been updated with the correct sequence, obtained from a new preparation of RNA from the hybridoma cells producing 4F2 mAb and partially confirmed by mass spectrometry. Although we currently have some new, unpublished evidences from mutagenesis studies indicating that the above mentioned conclusion is not absolutely wrong, we still
DOI of original article:10.1016/j.molimm.2006.03.004. ∗ Corresponding author. Fax: +53 7 2720644. E-mail address: [email protected] (A. López-Requena). 0161-5890/$ – see front matter © 2010 Elsevier Ltd. All rights reserved. doi:10.1016/j.molimm.2009.08.031
think that the right 4F2 mAb VL sequence sets an interesting scenario. The only differences between 1E10 and 4F2 mAbs reside in two amino acid residues, one at the heavy chain framework 1 (HFR1) (also confirmed by sequencing) and the other at the light chain CDR3 (L-CDR3); specifically, both the alanine 9 residue of VH and the glycine 91 residue of VL (Kabat numbering) in 1E10 (Pérez et al., 2001) are replaced by valine residues in 4F2, which additionally constitute mutations with respect to their most closely related VH and VL germ-line gene sequences reported in the Immunogenetics Database (IMGT, http://imgt.cines.fr). The change at 4F2 L-CDR3 seems to be enough for a number of differences between these antibodies: (i) a higher affinity of 4F2 for P3 mAb (Vázquez et al., 1998); (ii) no effect on the reactivity with 4F2 mAb of the two single and the double arginine-by-serine mutations at the heavy chain CDR3 (H-CDR3) of P3 mAb (López-Requena et al., 2007b), while the double mutation did affect in some extent the binding of 1E10 mAb (López-Requena et al., 2007a); (iii) the finding of 1E10-specific peptidic phagotopes that did not bind 4F2 mAb (Pérez et al., 2002; López-Requena et al., 2007b), suggesting that they were recognizing the 1E10 VL. In contrast, and also according to the model of the P3/1E10 (idiotype/anti-idiotype) complex obtained using the crystal structure of the chimeric P3 antibody Fab fragment (Talavera et al., Mol. Immunol., 2009), it is highly improbable that the amino acid replacement at H-FR1 could provide a satisfactory explanation for the differential binding properties of 1E10 and 4F2 mAbs. In fact, according to the model of the P3/1E10 complex, the L-CDR3 glycine 91 residue is tightly packed at the VH/VL interface; thus, the model predicts that a valine residue at that position would induce a conformational rearrangement of L-CDR3 in order to fit this bulkier amino acid. This rearrangement could lead to a better shape complementarity and a larger interacting surface between P3 and 4F2 mAbs, with respect to P3 and 1E10. Therefore, these data indicate that a single amino acid difference at L-CDR3 can have important implications for the antibody reactivity, even though other results suggest
940
A. López-Requena et al. / Molecular Immunology 47 (2010) 939–940
that in the binding to its antigen (in this case, another antibody) the heavy chain seems to have a preponderant role. We are currently attempting to crystallize the P3/1E10 complex to determine the definitive topology of this idiotype/anti-idiotype interaction. References López-Requena, A., Mateo de Acosta, C., Moreno, E., González, M., Puchades, Y., Talavera, A., Vispo, N.S., Vázquez, A.M., Pérez, R., 2007a. Gangliosides, Ab1 and Ab2 antibodies I. Towards a molecular dissection of an idiotype–anti-idiotype system. Mol. Immunol. 44, 423–433. López-Requena, A., Rodríguez, M., Mateo de Acosta, C., Moreno, E., Puchades, Y., González, M., Talavera, A., Valle, A., Hernández, T., Vázquez, A.M., Pérez, R., 2007b. Gangliosides, Ab1 and Ab2 antibodies II. Light versus heavy chain: an idiotype–anti-idiotype case study. Mol. Immunol. 44, 1015–1028. Pérez, A., Lombardero, J., Mateo, C., Mustelier, G., Alfonso, M., Vázquez, A.M., Pérez, R., 2001. Immunogenetic analysis of variable regions encoding Ab1 and ␥-type
Ab2 antibodies from the NeuGc-containing ganglioside family. Hybridoma 20, 211–221. Pérez, A., Mier, E.S., Vispo, N.S., Vázquez, A.M., Pérez, R., 2002. A monoclonal antibody against NeuGc-containing gangliosides contains a regulatory idiotope involved in the interaction with B and T cells. Mol. Immunol. 39, 103–112. Talavera, A., Eriksson, A., Okvist, M., López-Requena, A., Fernández-Marrero, Y., Pérez, R., Moreno, E., Krengel, U., 2009. Crystal structure of an anti-ganglioside antibody, and modelling of the functional mimicry of its NeuGc-GM3 antigen by an anti-idiotypic antibody. Mol. Immunol., doi:10.1016/j.molimm.2009.07.032. Vázquez, A.M., Alfonso, M., Lanne, B., Karlsson, K.-A., Carr, A., Barroso, O., Fernández, L.E., Rengifo, E., Lanio, M.E., Álvarez, C., Zeuthen, J., Pérez, R., 1995. Generation of a murine monoclonal antibody specific for N-glycolylneuraminic acid-containing gangliosides that also recognizes sulfated glycolipids. Hybridoma 14, 551– 556. Vázquez, A.M., Pérez, A., Hernández, A.M., Macías, A., Alfonso, M., Bombino, G., Pérez, R., 1998. Syngeneic anti-idiotypic monoclonal antibodies to an anti-NeuGccontaining ganglioside monoclonal antibody. Hybridoma 17, 527–534.