A complex between peptide:N-glycanase and two proteasome-linked proteins suggests a mechanism for the degradation of misfolded glycoproteins

被引:61
作者
Katiyar, S
Li, GT
Lennarz, WJ [1 ]
机构
[1] SUNY Stony Brook, Dept Biochem & Cell Biol, Stony Brook, NY 11794 USA
[2] SUNY Stony Brook, Inst Cell & Dev Biol, Stony Brook, NY 11794 USA
关键词
D O I
10.1073/pnas.0405663101
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
N-glycanase (PNGase) has been proposed to participate in the proteasome-dependent glycoprotein degradation pathway. The finding that yeast PNGase interacts with the 19S proteasome subunit through the protein Rad23 supports this hypothesis. In this report, we have used immunofluorescence, subcellular fractionation, coimmunoprecipitation, and in vitro GST pull-down techniques for detecting intracellular localization and interactions of PNGase, HR23B, and S4 by using human (h) and mouse (m) homologs. Immunofluorescence studies revealed that hPNGase, hHR23B, and hS4 are present in close proximity to the endoplasmic reticulum (ER) when calnexin was used as an ER marker in HeLa cells. Subcellular fractionation suggests not only cytoplasmic but also ER association of hPNGase in HeLa cells. Immunoprecipitation analysis revealed the interaction of h/mPNGase with the 19S proteasome subunit, hS4, through hHR23B. Using an in vitro GST pull-down assay, we also have shown that recombinant mPNGase requires its N terminus and middle domain for interaction with mHR23B. Finally, using misfolded yeast carboxypeptidase Y and chicken ovalbumin as glycoprotein substrates, we have established that mHR23B acts as a receptor for deglycosylated proteins. Based on this finding, we propose that after cleglycosylation of misfolded glycoproteins by PNGase, the aglyco forms of these proteins are recognized by HR23B and targeted for degradation.
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页码:13774 / 13779
页数:6
相关论文
共 45 条
  • [1] GTP-BINDING YPT1 PROTEIN AND CA-2+ FUNCTION INDEPENDENTLY IN A CELL-FREE PROTEIN-TRANSPORT REACTION
    BAKER, D
    WUESTEHUBE, L
    SCHEKMAN, R
    BOTSTEIN, D
    SEGEV, N
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1990, 87 (01) : 355 - 359
  • [2] UBA domains of DNA damage-inducible proteins interact with ubiquitin
    Bertolaet, BL
    Clarke, DJ
    Wolff, M
    Watson, MH
    Henze, M
    Divita, G
    Reed, SI
    [J]. NATURE STRUCTURAL BIOLOGY, 2001, 8 (05) : 417 - 422
  • [3] IMMUNOCYTOCHEMICAL LOCALIZATION OF THE MULTICATALYTIC PROTEINASE (PROTEASOME) IN CRUSTACEAN STRIATED MUSCLES
    BEYETTE, JR
    MYKLES, DL
    [J]. MUSCLE & NERVE, 1992, 15 (09) : 1023 - 1035
  • [4] A glycosylated type I membrane protein becomes cytosolic when peptide:: N-glycanase is compromised
    Blom, D
    Hirsch, C
    Stern, P
    Tortorella, D
    Ploegh, HL
    [J]. EMBO JOURNAL, 2004, 23 (03) : 650 - 658
  • [5] ER protein quality control and proteasome-mediated protein degradation
    Brodsky, JL
    McCracken, AA
    [J]. SEMINARS IN CELL & DEVELOPMENTAL BIOLOGY, 1999, 10 (05) : 507 - 513
  • [6] Ubiquitin-associated (UBA) domains in Rad23 bind ubiquitin and promote inhibition of multi-ubiquitin chain assembly
    Chen, L
    Shinde, U
    Ortolan, TG
    Madura, K
    [J]. EMBO REPORTS, 2001, 2 (10) : 933 - 938
  • [7] Rad23 promotes the targeting of proteolytic substrates to the proteasome
    Chen, L
    Madura, K
    [J]. MOLECULAR AND CELLULAR BIOLOGY, 2002, 22 (13) : 4902 - 4913
  • [8] CRESSWELL P, 1997, CURR BIOL, V7, P552
  • [9] Systematic identification of novel protein domain families associated with nuclear functions
    Doerks, T
    Copley, RR
    Schultz, J
    Ponting, CP
    Bork, P
    [J]. GENOME RESEARCH, 2002, 12 (01) : 47 - 56
  • [10] Proteasome subunit Rpn1 binds ubiquitin-like protein domains
    Elsasser, S
    Gali, RR
    Schwickart, M
    Larsen, CN
    Leggett, DS
    Müller, B
    Feng, MT
    Tübing, F
    Dittmar, GAG
    Finley, D
    [J]. NATURE CELL BIOLOGY, 2002, 4 (09) : 725 - 730