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PMID: 12815101 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Elucidation of the molecular logic by which misfolded alpha 1-antitrypsin is preferentially selected for degradation.

Wu Y, Swulius MT, Moremen KW, Sifers RN

Abstract

The exocytic pathway provides a physical route through which newly synthesized secretory and membrane proteins are deployed to the eukaryote cell surface. For newly synthesized alpha1-antitrypsin (AAT), the modification of its asparagine-linked oligosaccharides by a slow-acting mannosidase partitions the misfolded monomer into the proteasomal degradation pathway. Herein, we asked whether, and how, modification by endoplasmic reticulum mannosidase I (ERManI) contributes to the preferential selection of the misfolded AAT monomer for proteasomal degradation. Transiently expressed mutant and WT AAT variants underwent rapid destabilization in response to an artificially elevated ERManI concentration in the murine hepatoma cell line, Hepa1a. Based on the mannosidase- and lactacystin-sensitive properties of intracellular turnover, a stochastic model is proposed in which the delayed onset of the glycan modification, relative to the duration of nonnative protein structure, coordinates the preferential degradation of the misfolded monomer and spares the native molecule from destruction. Newly synthesized endogenous transferrin underwent degradation in response to an elevated concentration of ERManI, whereas the nonglycosylated secretory glycoprotein albumin was not affected. Taken together, these findings indicate that efficient conformational maturation might function as the initial quality control standard for a broad population of glycoproteins.

MeSH Terms
Acetylcysteine/analogs & derivatives,pharmacology Albumins/metabolism Amino Acid Sequence Animals Carbohydrate Sequence Endoplasmic Reticulum, Smooth/enzymology Enzyme Inhibitors/pharmacology Exocytosis Glycoproteins/chemistry,metabolism Glycosylation Liver Neoplasms, Experimental/pathology Mannans/metabolism Mannosidases/physiology Mice Models, Chemical Molecular Sequence Data Peptide Hydrolases/metabolism Proteasome Endopeptidase Complex Protein Conformation Protein Folding Protein Processing, Post-Translational Recombinant Fusion Proteins/physiology Stochastic Processes Structure-Activity Relationship Substrate Specificity Transfection Transferrin/metabolism Tumor Cells, Cultured/metabolism alpha 1-Antitrypsin/chemistry,genetics,metabolism
Chemicals
Albumins Enzyme Inhibitors Glycoproteins Mannans Recombinant Fusion Proteins Transferrin alpha 1-Antitrypsin mannosyl(9)-N-acetylglucosamine2 lactacystin Mannosidases mannosyl-oligosaccharide 1,2-alpha-mannosidase Peptide Hydrolases Proteasome Endopeptidase Complex ATP dependent 26S protease Acetylcysteine
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Wu Ying
Department of Pathology, Baylor College of Medicine, Houston, TX 77030-3498, USA.
Swulius Matthew T
Moremen Kelley W
Sifers Richard N
References (42)
42 references, click to expand
  1. A novel ER alpha-mannosidase-like protein accelerates ER-associated degradation.
    EMBO Rep. 2001 May;2(5):415-22 PMID: 11375934
  2. Alpha1-antitrypsin deficiency-associated liver disease progresses slowly in some children.
    J Pediatr Gastroenterol Nutr. 2000 Sep;31(3):258-63 PMID: 10997369
  3. Dissecting glycoprotein quality control in the secretory pathway.
    Trends Biochem Sci. 2001 Oct;26(10):619-24 PMID: 11590015
  4. The action of molecular chaperones in the early secretory pathway.
    Annu Rev Genet. 2001;35:149-91 PMID: 11700281
  5. The specificity of the yeast and human class I ER alpha 1,2-mannosidases involved in ER quality control is not as strict previously reported.
    Glycobiology. 2002 Apr;12(4):14G-15G PMID: 12090241
  6. E3 ubiquitin ligase that recognizes sugar chains.
    Nature. 2002 Jul 25;418(6896):438-42 PMID: 12140560
  7. Organizational diversity among distinct glycoprotein endoplasmic reticulum-associated degradation programs.
    Mol Biol Cell. 2002 Aug;13(8):2639-50 PMID: 12181335
  8. A time-dependent phase shift in the mammalian unfolded protein response.
    Dev Cell. 2003 Feb;4(2):265-71 PMID: 12586069
  9. Cell biology. Protein degradation unlocked.
    Science. 2003 Feb 28;299(5611):1330-1 PMID: 12610289
  10. EDEM as an acceptor of terminally misfolded glycoproteins released from calnexin.
    Science. 2003 Feb 28;299(5611):1394-7 PMID: 12610305
  11. Role of EDEM in the release of misfolded glycoproteins from the calnexin cycle.
    Science. 2003 Feb 28;299(5611):1397-400 PMID: 12610306
  12. Hepatoma secretory proteins migrate from rough endoplasmic reticulum to Golgi at characteristic rates.
    Nature. 1983 Jul 7-13;304(5921):80-3 PMID: 6866094
  13. Complete sequence of the cDNA for human alpha 1-antitrypsin and the gene for the S variant.
    Biochemistry. 1984 Oct 9;23(21):4828-37 PMID: 6093867
  14. A frameshift mutation results in a truncated alpha 1-antitrypsin that is retained within the rough endoplasmic reticulum.
    J Biol Chem. 1988 May 25;263(15):7330-5 PMID: 3259232
  15. A genetic shield to prevent emphysema?
    Science. 1989 Nov 10;246(4931):750-1 PMID: 2814494
  16. Intracellular degradation of the transport-impaired human PiZ alpha 1-antitrypsin variant. Biochemical mapping of the degradative event among compartments of the secretory pathway.
    J Biol Chem. 1990 Aug 15;265(23):14001-7 PMID: 2380201
  17. Accumulation of the insoluble PiZ variant of human alpha 1-antitrypsin within the hepatic endoplasmic reticulum does not elevate the steady-state level of grp78/BiP.
    J Biol Chem. 1990 Nov 25;265(33):20463-8 PMID: 2122976
  18. Glycosidase inhibitors: inhibitors of N-linked oligosaccharide processing.
    FASEB J. 1991 Dec;5(15):3055-63 PMID: 1743438
  19. Soluble aggregates of the human PiZ alpha 1-antitrypsin variant are degraded within the endoplasmic reticulum by a mechanism sensitive to inhibitors of protein synthesis.
    J Biol Chem. 1992 Jan 15;267(2):1072-80 PMID: 1530934
  20. Protein folding in the cell.
    Nature. 1992 Jan 2;355(6355):33-45 PMID: 1731198
  21. The mechanism of Z alpha 1-antitrypsin accumulation in the liver.
    Nature. 1992 Jun 18;357(6379):605-7 PMID: 1608473
  22. Molecular biology and genetics of alpha 1-antitrypsin deficiency.
    Semin Liver Dis. 1992 Aug;12(3):301-10 PMID: 1439881
  23. Conformational disease.
    Nat Cell Biol. 2000 Nov;2(11):E207-9 PMID: 11056553
  24. Pre-Golgi degradation of yeast prepro-alpha-factor expressed in a mammalian cell. Influence of cell type-specific oligosaccharide processing on intracellular fate.
    J Biol Chem. 1993 Jul 5;268(19):14301-9 PMID: 8314793
  25. Role of N-linked oligosaccharide recognition, glucose trimming, and calnexin in glycoprotein folding and quality control.
    Proc Natl Acad Sci U S A. 1994 Feb 1;91(3):913-7 PMID: 8302866
  26. Association between calnexin and a secretion-incompetent variant of human alpha 1-antitrypsin.
    J Biol Chem. 1994 Mar 11;269(10):7514-9 PMID: 8125971
  27. A lag in intracellular degradation of mutant alpha 1-antitrypsin correlates with the liver disease phenotype in homozygous PiZZ alpha 1-antitrypsin deficiency.
    Proc Natl Acad Sci U S A. 1994 Sep 13;91(19):9014-8 PMID: 8090762
  28. Inhibition of proteasome activities and subunit-specific amino-terminal threonine modification by lactacystin.
    Science. 1995 May 5;268(5211):726-31 PMID: 7732382
  29. What do dysfunctional serpins tell us about molecular mobility and disease?
    Nat Struct Biol. 1995 Feb;2(2):96-113 PMID: 7749926
  30. The Z type variation of human alpha 1-antitrypsin causes a protein folding defect.
    Nat Struct Biol. 1995 May;2(5):363-7 PMID: 7664092
  31. Defective protein folding as a basis of human disease.
    Trends Biochem Sci. 1995 Nov;20(11):456-9 PMID: 8578588
  32. Assembly of ER-associated protein degradation in vitro: dependence on cytosol, calnexin, and ATP.
    J Cell Biol. 1996 Feb;132(3):291-8 PMID: 8636208
  33. Intracellular disposal of incompletely folded human alpha1-antitrypsin involves release from calnexin and post-translational trimming of asparagine-linked oligosaccharides.
    J Biol Chem. 1997 Mar 21;272(12):7946-51 PMID: 9065464
  34. Endoplasmic reticulum degradation: reverse protein flow of no return.
    FASEB J. 1997 Dec;11(14):1227-33 PMID: 9409541
  35. Degradation of misfolded endoplasmic reticulum glycoproteins in Saccharomyces cerevisiae is determined by a specific oligosaccharide structure.
    J Cell Biol. 1998 Sep 7;142(5):1223-33 PMID: 9732283
  36. Ubiquitin and the control of protein fate in the secretory and endocytic pathways.
    Annu Rev Cell Dev Biol. 1998;14:19-57 PMID: 9891777
  37. Oligosaccharide modification in the early secretory pathway directs the selection of a misfolded glycoprotein for degradation by the proteasome.
    J Biol Chem. 1999 Feb 26;274(9):5861-7 PMID: 10026209
  38. Identification, expression, and characterization of a cDNA encoding human endoplasmic reticulum mannosidase I, the enzyme that catalyzes the first mannose trimming step in mammalian Asn-linked oligosaccharide biosynthesis.
    J Biol Chem. 1999 Jul 23;274(30):21375-86 PMID: 10409699
  39. Cloning and expression of a specific human alpha 1,2-mannosidase that trims Man9GlcNAc2 to Man8GlcNAc2 isomer B during N-glycan biosynthesis.
    Glycobiology. 1999 Oct;9(10):1073-8 PMID: 10521544
  40. Setting the standards: quality control in the secretory pathway.
    Science. 1999 Dec 3;286(5446):1882-8 PMID: 10583943
  41. Processing by endoplasmic reticulum mannosidases partitions a secretion-impaired glycoprotein into distinct disposal pathways.
    J Biol Chem. 2000 Aug 11;275(32):25015-22 PMID: 10827201
  42. ER quality control: towards an understanding at the molecular level.
    Curr Opin Cell Biol. 2001 Aug;13(4):431-7 PMID: 11454449
Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
Published
2003-07-08
Epub
2003-00-18
Pages
8229-34
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC166211
Subset
IM
Grants
NIGMS NIH HHS · GM47533 · United States
NCRR NIH HHS · P41 RR005351 · United States
NCRR NIH HHS · RR05351 · United States
NIGMS NIH HHS · R01 GM047533 · United States
NHLBI NIH HHS · HL62553 · United States
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