Endoplasmic reticulum quality control of asialoglycoprotein receptor H2a involves a determinant for retention and not retrieval.
The human asialoglycoprotein receptor H2a subunit contains a charged pentapeptide, EGHRG, in its ectodomain that is the only sequence absent from the H2b alternatively spliced variant. H2b exits the endoplasmic reticulum (ER) even when singly expressed, whereas H2a gives rise to a cleaved soluble secreted ectodomain fragment; uncleaved membrane-bound H2a molecules are completely retained and degraded in the ER. We have inserted the H2a pentapeptide into the sequence of the H1 subunit (H1i5), which caused complete ER retention but, unexpectedly, no degradation. This suggests that the pentapeptide is a determinant for ER retention not colocalizing in H2a with the determinant for degradation. The state of sugar chain processing and the ER localization of H1i5, which was unchanged at 15 degrees C or after treatment with nocodazole, indicate ER retention and not retrieval from the cis-Golgi or the intermediate compartment. H1i5 folded similarly to H1, and both associated to calnexin. However, whereas H1 dissociated with a half time of 45 min, H1i5 remained bound to the chaperone for prolonged periods. The correct global folding of H2a and H1i5 and of other normal precursors and unassembled proteins and the true ER retention, and not exit and retrieval, suggest a difference in their quality control mechanism compared with that of misfolded proteins, which does involve retrieval. However, both pathways may involve calnexin.
Shenkman
M
M
Department of Cell Research and Immunology, George Wise Faculty of Life Sciences, Tel Aviv University, Tel Aviv, Israel, 69978, USA.
Ayalon
M
M
Lederkremer
G Z
GZ
D016475:3T3 Cells; D000595:Amino Acid Sequence; D000818:Animals; D037263:Asialoglycoprotein Receptor; D020126:Brefeldin A; D002135:Calcium-Binding Proteins / Q000502:physiology; D037281:Calnexin; D002240:Carbohydrate Sequence; D002462:Cell Membrane / Q000502:physiology; D003517:Cyclopentanes / Q000494:pharmacology; D004721:Endoplasmic Reticulum / Q000187:drug effects / Q000502:physiology*; D006056:Golgi Apparatus / Q000502:physiology; D006801:Humans; D046911:Macromolecular Substances; D051379:Mice; D018832:Molecular Chaperones / Q000502:physiology; D008969:Molecular Sequence Data; D015739:Nocodazole / Q000494:pharmacology; D010446:Peptide Fragments; D017510:Protein Folding; D011956:Receptors, Cell Surface / Q000737:chemistry / Q000502:physiology*; D011994:Recombinant Proteins / Q000378:metabolism; D016415:Sequence Alignment; D017026:Swainsonine / Q000494:pharmacology; D014162:Transfection
D037263:Asialoglycoprotein Receptor; D002135:Calcium-Binding Proteins; D003517:Cyclopentanes; D046911:Macromolecular Substances; D018832:Molecular Chaperones; D010446:Peptide Fragments; D011956:Receptors, Cell Surface; D011994:Recombinant Proteins; D037281:Calnexin; D020126:Brefeldin A; D017026:Swainsonine; D015739:Nocodazole