HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Andres, D. A. Articles by Dixon, J. E. Search for Related Content PubMed PubMed Citation Articles by Andres, D. A. Articles by Dixon, J. E. Social Bookmarking                      What's this?

J. Biol. Chem., Vol. 265, Issue 11, 5952-5955, Apr, 1990

Variants of the carboxyl-terminal KDEL sequence direct intracellular retention

DA Andres, IM Dickerson and JE Dixon
Department of Biochemistry, Purdue University, West Lafayette, Indiana 47907.

Soluble proteins which reside in the lumen of the endoplasmic reticulum share a common carboxyl-terminal tetrapeptide Lys-Asp-Glu-Leu (KDEL). Addition of the tetrapeptide to a normally secreted protein is both necessary and sufficient to cause retention in the endoplasmic reticulum. In order to characterize the critical residues in the KDEL signal, cDNAs encoding proneuropeptide Y (pro-NPY) with the 4-amino acid carboxyl-terminal extension KDEL or a series of KDEL variants were expressed in the AtT-20 cell line. AtT-20 cells, a mouse anterior pituitary corticotrope cell line, synthesize, process, and secrete the pro-ACTH/endorphin precursor. Since post-translational processing in AtT-20 cells has been extensively characterized, it provides a model system in which the processing of a foreign peptide precursor (pro-NPY) and the endogenous precursor (pro-ACTH/endorphin) can be compared. Altered cDNAs encoding pro-NPY with KDEL, DKEL, RDEL, KNEL, KDQL, or KDEA at the COOH terminus were used to generate stable AtT-20 cell lines. The processing of pro-NPY to neuropeptide Y and the carboxyl- terminal peptide was studied using sodium dodecyl sulfate- polyacrylamide gel electrophoresis, tryptic peptide mapping, and radiosequencing. Addition of the tetrapeptides KDEL, DKEL, RDEL, or KNEL to the COOH terminus of the neuropeptide Y precursor, a peptide hormone normally processed and secreted from neuronal cells, caused complete intracellular retention of the unprocessed prohormone in AtT- 20 cells. However, KDQL and KDEA-extended pro-NPY molecules were processed and secreted like wild-type pro-NPY when expressed in AtT-20 cells. The secretion of proNPY-derived peptides in these cell lines paralleled secretion of endogenous pro-ACTH/endorphin-derived products under both basal and stimulated conditions. These mutagenesis studies demonstrate that variants of the KDEL retention signal can direct intracellular retention.
CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				S. C. P. Eschenlauer and A. P. Page The Caenorhabditis elegans ERp60 Homolog Protein Disulfide Isomerase-3 Has Disulfide Isomerase and Transglutaminase-like Cross-linking Activity and Is Involved in the Maintenance of Body Morphology J. Biol. Chem., January 31, 2003; 278(6): 4227 - 4237. [Abstract] [Full Text] [PDF]

				T. Wex, F. Buhling, H. Wex, D. Gunther, P. Malfertheiner, E. Weber, and D. Bromme Human Cathepsin W, a Cysteine Protease Predominantly Expressed in NK Cells, Is Mainly Localized in the Endoplasmic Reticulum J. Immunol., August 15, 2001; 167(4): 2172 - 2178. [Abstract] [Full Text] [PDF]

				L. J.-s. Huang, L. Wang, Y. Ma, K. Durick, G. Perkins, T. J. Deerinck, M. H. Ellisman, and S. S. Taylor NH2-Terminal Targeting Motifs Direct Dual Specificity A-Kinase-anchoring Protein 1 (D-AKAP1) to Either Mitochondria or Endoplasmic Reticulum J. Cell Biol., May 31, 1999; 145(5): 951 - 959. [Abstract] [Full Text] [PDF]

				D. Yabe, T. Nakamura, N. Kanazawa, K. Tashiro, and T. Honjo Calumenin, a Ca2+-binding Protein Retained in the Endoplasmic Reticulum with a Novel Carboxyl-terminal Sequence, HDEF J. Biol. Chem., July 18, 1997; 272(29): 18232 - 18239. [Abstract] [Full Text] [PDF]

				G Bu, S Rennke, and H. Geuze ERD2 proteins mediate ER retention of the HNEL signal of LRP's receptor-associated protein (RAP) J. Cell Sci., January 1, 1997; 110(1): 65 - 73. [Abstract] [PDF]

				B Sonnichsen, J Fullekrug, P Nguyen Van, W Diekmann, D. Robinson, and G Mieskes Retention and retrieval: both mechanisms cooperate to maintain calreticulin in the endoplasmic reticulum J. Cell Sci., January 10, 1994; 107(10): 2705 - 2717. [Abstract] [PDF]