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.
引用
收藏
页码:13774 / 13779
页数:6
相关论文
共 45 条
  • [11] Subcellular distribution of proteasomes implicates a major location of protein degradation in the nuclear envelope ER network in yeast
    Enenkel, C
    Lehmann, A
    Kloetzel, PM
    [J]. EMBO JOURNAL, 1998, 17 (21) : 6144 - 6154
  • [12] GFP-labelling of 26S proteasomes in living yeast: insight into proteasomal functions at the nuclear envelope rough ER
    Enenkel, C
    Lehmann, A
    Kloetzel, PM
    [J]. MOLECULAR BIOLOGY REPORTS, 1999, 26 (1-2) : 131 - 135
  • [13] Subunit interaction maps for the regulatory particle of the 26S proteasome and the COP9 signalosome
    Fu, HY
    Reis, N
    Lee, Y
    Glickman, MH
    Vierstra, RD
    [J]. EMBO JOURNAL, 2001, 20 (24) : 7096 - 7107
  • [14] Quaternary structure of the ATPase complex of human 26S proteasomes determined by chemical cross-linking
    Hartmann-Petersen, R
    Tanaka, K
    Hendil, KB
    [J]. ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 2001, 386 (01) : 89 - 94
  • [15] A role for N-glycanase in the cytosolic turnover of glycoproteins
    Hirsch, C
    Blom, D
    Ploegh, HL
    [J]. EMBO JOURNAL, 2003, 22 (05) : 1036 - 1046
  • [16] Interaction of hHR23 with S5a - The ubiquitin-like domain of hHR23 mediates interaction with S5a subunit of 26 S proteasome
    Hiyama, H
    Yokoi, M
    Masutani, C
    Sugasawa, K
    Maekawa, T
    Tanaka, K
    Hoeijmakers, JHJ
    Hanaoka, F
    [J]. JOURNAL OF BIOLOGICAL CHEMISTRY, 1999, 274 (39) : 28019 - 28025
  • [17] A novel quality control compartment derived from the endoplasmic reticulum
    Kamhi-Nesher, S
    Shenkman, M
    Tolchinsky, S
    Fromm, SV
    Ehrlich, R
    Lederkremer, GZ
    [J]. MOLECULAR BIOLOGY OF THE CELL, 2001, 12 (06) : 1711 - 1723
  • [18] Site-directed mutagenesis study of yeast peptide:: N-glycanase -: Insight into the reaction mechanism of deglycosylation
    Katiyar, S
    Suzuki, T
    Balgobin, BJ
    Lennarz, WJ
    [J]. JOURNAL OF BIOLOGICAL CHEMISTRY, 2002, 277 (15) : 12953 - 12959
  • [19] Processing of N-linked glycans during endoplasmic-reticulum-associated degradation of a short-lived variant of ribophorin I
    Kitzmüller, C
    Caprini, A
    Moore, SEH
    Frénoy, JP
    Schwaiger, E
    Kellermann, O
    Ivessa, NE
    Ermonval, M
    [J]. BIOCHEMICAL JOURNAL, 2003, 376 : 687 - 696
  • [20] The axial channel of the proteasome core particle is gated by the Rpt2 ATPase and controls both substrate entry and product release
    Köhler, A
    Cascio, P
    Leggett, DS
    Woo, KM
    Goldberg, AL
    Finley, D
    [J]. MOLECULAR CELL, 2001, 7 (06) : 1143 - 1152