Home | Help | Feedback | Subscriptions | Archive | Search | Table of Contents

This Article Full Text Full Text (PDF, 1852K) PPT slides of all figures Alert me when this article is cited Citation Map Services Email this article Similar articles in this journal Similar articles in PubMed Alert me to new content in the JEM Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Huppa, J. B. Articles by Ploegh, H. L. Search for Related Content PubMed PubMed Citation Articles by Huppa, J. B. Articles by Ploegh, H. L. Social Bookmarking                            What's this?

Articles

From the Center for Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139

The T cell receptor for antigen (TCR) is a multisubunit complex that consists of at least seven polypeptides: the clonotypic, disulfide-linked /β heterodimer that is noncovalently associated with the invariant polypeptides of the CD3 complex (CD3-, -, -) and , a disulfide-linked homodimer. We achieved the complete assembly of the human TCR in an in vitro transcription/translation system supplemented with dog pancreas microsomes by simultaneous translation of the messenger RNAs encoding the TCR-, -β and CD3-, -, -, and - subunits. CD3-, one of the subunits that initiates the assembly of the TCR in living cells, forms misfolded, disulfide-linked homooligomers when translated alone. However, co-translation of one of its first binding partners in the course of assembly, CD3- or -, led to the expression of mainly monomeric and correctly folded subunits, the only form we could detect as part of a properly assembled TCR complex. In the absence of these subunits, the ER-resident chaperone calnexin interacted with oligomeric, i.e. misfolded, structures of CD3- in a glycan-independent manner. A glycan-dependent interaction between CD3- and calnexin was mediated by CD3- and concerned only monomeric CD3- complexed with CD3-, but was dispensable for proper folding of CD3-. We suggest that in addition to its signaling function, CD3- serves as a monitor for proper subunit assembly of the TCR.

Johannes B. Huppa was supported by fellowship of the Daimler-Benz-Stiftung (Ladenburg, Germany) and is currently a fellow of the Boehringer-Ingelheim-Fonds (Stuttgart, Germany). This work received support from the National Institutes of Health (1 PO1 AI 37833-01).

¹ Abbreviations used in this paper: 7-O-dec, N-7-oxadecyl-dNM; DTT, dithiothreitol; EndoH, endoglycosidase H; ER, endoplasmic reticulum; GSSG, oxidized glutathion; HA, hemagglutinin; mRNA, messenger RNA; VSV–G protein, vesicular stomatitis virus G protein.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Facebook    Reddit    Technorati    Twitter    What's this?

This article has been cited by other articles:

• Xu, C., Call, M. E., Wucherpfennig, K. W. (2006). A Membrane-proximal Tetracysteine Motif Contributes to Assembly of CD3{delta}{epsilon} and CD3{gamma}{epsilon} Dimers with the T Cell Receptor. J. Biol. Chem. 281: 36977-36984 [Abstract] [Full Text]  
• Mueller, B., Lilley, B. N., Ploegh, H. L. (2006). SEL1L, the homologue of yeast Hrd3p, is involved in protein dislocation from the mammalian ER. JCB 175: 261-270 [Abstract] [Full Text]  
• Taylor, K. N., Shinde Patil, V. R., Colson, Y. L. (2006). Reconstitution of Allogeneic Hemopoietic Stem Cells: The Essential Role of FcR{gamma} and the TCR beta-Chain-FCp33 Complex. J. Immunol. 177: 1444-1450 [Abstract] [Full Text]  
• Wang, N., Daniels, R., Hebert, D. N. (2005). The Cotranslational Maturation of the Type I Membrane Glycoprotein Tyrosinase: The Heat Shock Protein 70 System Hands Off to the Lectin-based Chaperone System. Mol. Biol. Cell 16: 3740-3752 [Abstract] [Full Text]  
• Garrity, D., Call, M. E., Feng, J., Wucherpfennig, K. W. (2005). The activating NKG2D receptor assembles in the membrane with two signaling dimers into a hexameric structure. Proc. Natl. Acad. Sci. USA 102: 7641-7646 [Abstract] [Full Text]  
• Ruder, C., Reimer, T., Delgado-Martinez, I., Hermosilla, R., Engelsberg, A., Nehring, R., Dorken, B., Rehm, A. (2005). EBAG9 Adds a New Layer of Control on Large Dense-Core Vesicle Exocytosis via Interaction with Snapin. Mol. Biol. Cell 16: 1245-1257 [Abstract] [Full Text]  
• Fiebiger, E., Tortorella, D., Jouvin, M.-H., Kinet, J.-P., Ploegh, H. L. (2005). Cotranslational endoplasmic reticulum assembly of Fc{varepsilon}RI controls the formation of functional IgE-binding receptors. JEM 201: 267-277 [Abstract] [Full Text]  
• Esapa, C. T., McIlhinney, R.A. J., Blake, D. J. (2005). Fukutin-related protein mutations that cause congenital muscular dystrophy result in ER-retention of the mutant protein in cultured cells. Hum Mol Genet 14: 295-305 [Abstract] [Full Text]  
• Kjer-Nielsen, L., Dunstone, M. A., Kostenko, L., Ely, L. K., Beddoe, T., Mifsud, N. A., Purcell, A. W., Brooks, A. G., McCluskey, J., Rossjohn, J. (2004). Crystal structure of the human T cell receptor CD3{epsilon}{gamma} heterodimer complexed to the therapeutic mAb OKT3. Proc. Natl. Acad. Sci. USA 101: 7675-7680 [Abstract] [Full Text]  
• Fiebiger, E., Hirsch, C., Vyas, J. M., Gordon, E., Ploegh, H. L., Tortorella, D. (2004). Dissection of the Dislocation Pathway for Type I Membrane Proteins with a New Small Molecule Inhibitor, Eeyarestatin. Mol. Biol. Cell 15: 1635-1646 [Abstract] [Full Text]  
• Ghadessy, F. J., Holliger, P. (2004). A novel emulsion mixture for in vitro compartmentalization of transcription and translation in the rabbit reticulocyte system. Protein Eng Des Sel 17: 201-204 [Abstract] [Full Text]  
• Furman, M. H., Loureiro, J., Ploegh, H. L., Tortorella, D. (2003). Ubiquitinylation of the Cytosolic Domain of a Type I Membrane Protein Is Not Required to Initiate Its Dislocation from the Endoplasmic Reticulum. J. Biol. Chem. 278: 34804-34811 [Abstract] [Full Text]  
• Kosolapov, A., Deutsch, C. (2003). Folding of the Voltage-gated K+ Channel T1 Recognition Domain. J. Biol. Chem. 278: 4305-4313 [Abstract] [Full Text]  
• Rehm, A., Engelsberg, A., Tortorella, D., Korner, I. J., Lehmann, I., Ploegh, H. L., Hopken, U. E. (2002). Human Cytomegalovirus Gene Products US2 and US11 Differ in Their Ability To Attack Major Histocompatibility Class I Heavy Chains in Dendritic Cells. J. Virol. 76: 5043-5050 [Abstract] [Full Text]  
• Law, C.-L., Hayden-Ledbetter, M., Buckwalter, S., McNeill, L., Nguyen, H., Habecker, P., Thorne, B. A., Dua, R., Ledbetter, J. A. (2002). Expression and characterization of recombinant soluble human CD3 molecules: presentation of antigenic epitopes defined on the native TCR-CD3 complex. Int Immunol 14: 389-400 [Abstract] [Full Text]  
• Gewurz, B. E., Ploegh, H. L., Tortorella, D. (2002). US2, a Human Cytomegalovirus-encoded Type I Membrane Protein, Contains a Non-cleavable Amino-terminal Signal Peptide. J. Biol. Chem. 277: 11306-11313 [Abstract] [Full Text]  
• Merola, M., Brazzoli, M., Cocchiarella, F., Heile, J. M., Helenius, A., Weiner, A. J., Houghton, M., Abrignani, S. (2001). Folding of Hepatitis C Virus E1 Glycoprotein in a Cell-Free System. J. Virol. 75: 11205-11217 [Abstract] [Full Text]  
• Kirchgessner, H., Dietrich, J., Scherer, J., Isomaki, P., Korinek, V., Hilgert, I., Bruyns, E., Leo, A., Cope, A. P., Schraven, B. (2001). The Transmembrane Adaptor Protein Trim Regulates T Cell Receptor (Tcr) Expression and Tcr-Mediated Signaling via an Association with the Tcr {zeta} Chain. JEM 193: 1269-1284 [Abstract] [Full Text]  
• Petersson, K., Ivars, F. (2001). Early TCR {{alpha}}{{beta}} Expression Promotes Maturation of T Cells Expressing Fc{{epsilon}}RI{{gamma}} Containing TCR/CD3 Complexes. J. Immunol. 166: 6616-6624 [Abstract] [Full Text]  
• Alibaud, L., Llobera, R., Al Saati, T., March, M., Delsol, G., Rubin, B. (2000). A New Monoclonal Anti-CD3{varepsilon} Antibody Reactive on Paraffin Sections. J. Histochem. Cytochem. 48: 1609-1616 [Abstract] [Full Text]  
• DeLeo, F. R., Burritt, J. B., Yu, L., Jesaitis, A. J., Dinauer, M. C., Nauseef, W. M. (2000). Processing and Maturation of Flavocytochrome b558 Include Incorporation of Heme as a Prerequisite for Heterodimer Assembly. J. Biol. Chem. 275: 13986-13993 [Abstract] [Full Text]  
• Gobel, T. W. F., Dangy, J.-P. (2000). Evidence for a Stepwise Evolution of the CD3 Family. J. Immunol. 164: 879-883 [Abstract] [Full Text]  
• Appel, H., Gauthier, L., Pyrdol, J., Wucherpfennig, K. W. (2000). Kinetics of T-cell Receptor Binding by Bivalent HLA-DR{middle dot}Peptide Complexes That Activate Antigen-specific Human T-cells. J. Biol. Chem. 275: 312-321 [Abstract] [Full Text]  
• Nolan, K. F., Yun, C.-O., Akamatsu, Y., Murphy, J. C., Leung, S.-O., Beecham, E. J., Junghans, R. P. (1999). Bypassing Immunization: Optimized Design of ""Designer T Cells"" against Carcinoembryonic Antigen (CEA)-expressing Tumors, and Lack of Suppression by Soluble CEA. Clin. Cancer Res. 5: 3928-3941 [Abstract] [Full Text]  
• Bronstein-Sitton, N., Wang, L., Cohen, L., Baniyash, M. (1999). Expression of the T Cell Antigen Receptor zeta Chain following Activation Is Controlled at Distinct Checkpoints. IMPLICATIONS FOR CELL SURFACE RECEPTOR DOWN-MODULATION AND RE-EXPRESSION. J. Biol. Chem. 274: 23659-23665 [Abstract] [Full Text]  
• Borroto, A., Lama, J., Niedergang, F., Dautry-Varsat, A., Alarcon, B., Alcover, A. (1999). The CD3{epsilon} Subunit of the TCR Contains Endocytosis Signals. J. Immunol. 163: 25-31 [Abstract] [Full Text]  
• Ohsako, S., Janulis, L., Hayashi, Y., Bunick, D. (1998). Characterization of Domains in Mice of Calnexin-t, a Putative Molecular Chaperone Required in Sperm Fertility, with Use of Glutathione S-Transferase-Fusion Proteins. Biol. Reprod. 59: 1214-1223 [Abstract] [Full Text]  
• Schust, D. J., Tortorella, D., Seebach, J., Phan, C., Ploegh, H. L. (1998). Trophoblast Class I Major Histocompatibility Complex (MHC) Products Are Resistant to Rapid Degradation Imposed by the Human Cytomegalovirus (HCMV) Gene Products US2 and US11. JEM 188: 497-503 [Abstract] [Full Text]  
• Kearse, K. P. (1998). Calnexin Associates with Monomeric and Oligomeric (Disulfide-linked) CD3delta Proteins in Murine T Lymphocytes. J. Biol. Chem. 273: 14152-14157 [Abstract] [Full Text]  
• Ghendler, Y., Smolyar, A., Chang, H.-C., Reinherz, E. L. (1998). One of the CD3{varepsilon} Subunits within a T Cell Receptor Complex Lies in Close Proximity to the C{beta} FG Loop. JEM 187: 1529-1536 [Abstract] [Full Text]  
• Ujvari, A., Aron, R., Eisenhaure, T., Cheng, E., Parag, H. A., Smicun, Y., Halaban, R., Hebert, D. N. (2001). Translation Rate of Human Tyrosinase Determines Its N-Linked Glycosylation Level. J. Biol. Chem. 276: 5924-5931 [Abstract] [Full Text]  
• Furman, M. H., Ploegh, H. L., Tortorella, D. (2002). Membrane-specific, Host-derived Factors Are Required for US2- and US11-mediated Degradation of Major Histocompatibility Complex Class I Molecules. J. Biol. Chem. 277: 3258-3267 [Abstract] [Full Text]  

Home | Help | Feedback | Subscriptions | Archive | Search

TABLE OF CONTENTS