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Interaction the CDR2-Containing Region for Igm in the Polymeric Ig

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Interaction the CDR2-Containing Region for Igm in the Polymeric Ig Fine Specificity of Ligand-Binding Domain 1 in the Polymeric Ig Receptor: Importance of the CDR2-Containing Region for IgM Interaction This information is current as of September 24, 2021. Målfrid Røe, Inger N. Norderhaug, Per Brandtzaeg and Finn-Eirik Johansen J Immunol 1999; 162:6046-6052; ; http://www.jimmunol.org/content/162/10/6046 Downloaded from References This article cites 50 articles, 22 of which you can access for free at: http://www.jimmunol.org/content/162/10/6046.full#ref-list-1 http://www.jimmunol.org/ Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists • Fast Publication! 4 weeks from acceptance to publication by guest on September 24, 2021 *average Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 1999 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. Fine Specificity of Ligand-Binding Domain 1 in the Polymeric Ig Receptor: Importance of the CDR2-Containing Region for IgM Interaction1 Målfrid Røe, Inger N. Norderhaug, Per Brandtzaeg, and Finn-Eirik Johansen2 The human polymeric Ig receptor (pIgR), also called transmembrane secretory component, is expressed basolaterally on exocrine epithelia, and mediates specific external transport of dimeric IgA and pentameric IgM. The extracellular part of pIgR consists of five Ig-like domains (D1-D5), and a highly conserved D1 region appears to mediate the initial noncovalent ligand interaction. While the human pIgR binds both dimeric IgA and pentameric IgM with high affinity, the rabbit counterpart has virtually no binding capacity for pentameric IgM. This remarkable disparity constitutes evidence that the binding site of the two ligands differs with regard to essential receptor contact elements. Therefore, we expressed human/rabbit chimeric pIgRs in Madin-Darby canine Downloaded from kidney cells and found that human pIgR D1 is crucial for the interaction with pentameric IgM when placed in the context of a full-length receptor regardless of its backbone species. D1 contains three complementarity-determining region-like loops (CDR1– 3), and to further map human D1 regions involved in pentameric IgM binding, we transfected Madin-Darby canine kidney cells with human/rabbit chimeric receptors in which the regions containing the CDR-like loops had been interchanged. Our results showed that the region containing the CDR2-like loop is the most essential for pentameric IgM binding. The region containing the CDR1-like loop also contributed substantially to this interaction, whereas only little contribution was provided by the region http://www.jimmunol.org/ containing the CDR3-like loop, although it appeared to be necessary for maximal pentameric IgM binding. The Journal of Immunology, 1999, 162: 6046–6052. ecretory Ig (SIgA3 and SIgM) Abs play a major role in for pIgR in both pIgA and pentameric IgM, it is unknown how the adaptive defense at mucosal surfaces, the largest body area polymers themselves are involved in this binding site. The fact that S exposed to the external environment (1–4). Dimers and pentameric IgM has been shown to interact with free SC with an larger polymers of IgA (collectively called pIgA) and pentameric affinity that is 8–30 times that determined for pIgA (16, 17) sug- IgM synthesized by subepithelial plasma cells become specifically gests that the pIgR binding site of the two polymers is structurally bound by the human pIg receptor (pIgR), also known as the trans- somewhat different. This idea is supported by remarkable species by guest on September 24, 2021 membrane secretory component (SC). This receptor is expressed differences shown by pIgR with regard to pentameric IgM inter- on the basolateral surface of secretory epithelial cells (5, 6). The action (see below). pIgs are next internalized and transported across these cells to their The pIgR is a glycoprotein of 100–120 kDa (depending on the apical domain, where the extracellular ligand-binding portion of species) with five Ig-like extracellular domains (D1-D5) that are the receptor is proteolytically cleaved and released to the lumen, structurally most similar to the IgV regions (18). Binding of pIgA either complexed to the ligand as bound SC or unoccupied as free to pIgR appears to be a sequential process in which an initial SC (4, 7). noncovalent ligand interaction with D1 progresses to other do- It remains an enigma that two so structurally different polymers mains (Ref. 19 and Norderhaug et al.4) and is followed (in most as pIgA and pentameric IgM can bind specifically to the same species) by disulfide binding between one of the IgA heavy chains receptor, although their shared J chain has been shown to be es- and D5 (reviewed in Ref. 20). Several lines of evidence suggest sential for this interaction (8–15). However, by itself this polypep- that D1 carries the primary site of interaction with pIgA. First, both tide shows only marginal affinity for free SC (10). Therefore, al- proteolytic and recombinant fragments of the receptor that contain though the J chain in a crucial way contributes to the binding site D1 have been shown to retain the capacity to bind pIgA, although this initial interaction with certain synthetic receptor peptides ap- pears to be “promiscuous” with regard to Ig class (19, 21–23) (see Laboratory for Immunohistochemistry and Immunopathology (LIIPAT), Institute of Pathology, University of Oslo, The National Hospital, Rikshospitalet, Oslo, Norway below). Second, mAbs that recognize an epitope within D1 have Received for publication November 16, 1998. Accepted for publication March been shown to compete with pIgA for binding to the receptor (21). 3, 1999. Furthermore, we have shown recently that binding of pentameric The costs of publication of this article were defrayed in part by the payment of page IgM to the human pIgR depends preferentially on a strong inter- charges. This article must therefore be hereby marked advertisement in accordance action with D1, while binding of pIgA in addition depends on with 18 U.S.C. Section 1734 solely to indicate this fact. determinants within D2 and/or D3 to support the initial noncova- 1 This study was supported by the University of Oslo, the Research Council of Nor- lent interaction with D1.4 way, the Norwegian Cancer Society, and Anders Jahre’s Foundation for Promotion of Science. M.R. has been a Research Fellow of the University of Oslo. Located in D1 are three loops corresponding to the complemen- 2 Address correspondence and reprint requests to Dr. Finn-Eirik Johansen, LIIPAT, tarity-determining regions (CDR1-CDR3) of IgV regions (24), the Institute of Pathology, Rikshospitalet, N-0027 Oslo, Norway. E-mail address: [email protected] 3 Abbreviations used in this paper: SIg, secretory Ig; CDR, complementarity-deter- 4 I. N. Norderhaug, F.-E. Johansen, P. Krajci, and P. Brandtzaeg. Domain deletions in mining region; MDCK, Madin-Darby canine kidney; pIgA, polymeric IgA; pIgR, the human polymeric Ig receptor disclose differences between its dimeric IgA and polymeric IgR, SC, secretory component. pentameric IgM interaction. Submitted for publication. Copyright © 1999 by The American Association of Immunologists 0022-1767/99/$02.00 The Journal of Immunology 6047 sequence that determines their Ag-binding specificity. The CDR1- like loop in pIgR D1 is highly conserved among different species, 82–100% when conservative amino acid changes are not taken into account (25). The CDR2- and CDR3-like loops in D1 show less interspecies homology, but retain some invariant residues that may play important roles in ligand binding. Thus, a study by Coyne et al. (25), based on a mutational approach with modeling of the rabbit pIgR D1 sequence on known Ig variable structures, sug- gested that all three loops participate in the specific noncovalent binding of human pIgA. The order of importance was not exam- ined, but based on the studies mentioned above, the CDR1-like loop is probably the most important one for the initial interaction. In humans, pIgR binding of pIgA also depends on structural ele- ments outside D1 (Ref. 19 and Norderhaug et al.4), and the im- portance of interactions between pIgA and elements in CDR2 and CDR3 may therefore be less significant than for rabbit D1. Human pIgR does not show stable binding of monomeric IgA or other monomeric Ig isotypes, but interacts with both pIgA and pentameric IgM with high affinity, as mentioned above (16, 26). In Downloaded from mice and rabbits, on the other hand, the pIgR binds primarily pIgA (27). To characterize the interaction of human pentameric IgM with pIgR, we exploited this species difference and generated sev- eral chimeric receptors to characterize the pentameric IgM binding site of the receptor. We found that D1 of human origin could transfer its pentameric IgM-binding affinity to the rabbit pIgR, thus http://www.jimmunol.org/ substantiating its crucial role for both pIgA and pentameric IgM interaction. Furthermore, we demonstrated that the D1 regions containing CDR1- and CDR2-like loops of human origin could transfer substantial pentameric IgM-binding capacity to the rabbit pIgR. However, all three human pIgR CDR-like regions were re- quired for maximal pentameric IgM-binding capacity. Materials and Methods Immunoglobulins FIGURE 1. A, Comparison of the deduced amino acid sequences of the by guest on September 24, 2021 experimentally interchanged regions A, B, and C of human (H) and rabbit Polyclonal human pIgA (28), mainly IgA1, crude monomeric IgA, and (R) pIgR domain 1 (upper and lower lines, respectively).
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