Cited by 86Ubiquitin and proteasome pathwaysGlycosylation and Glycoproteins ResearchEndoplasmic Reticulum Stress and Disease

Diversity in Tissue Expression, Substrate Binding, and SCF Complex Formation for a Lectin Family of Ubiquitin Ligases

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Glenn, Kevin A., Nelson, Rick F., Wen, Hsiang M., Mallinger, Adam J., Paulson, Henry L.·2008·Journal of Biological Chemistry

Abstract

Post-translational modification of proteins regulates many cellular processes. Some modifications, including N-linked glycosylation, serve multiple functions. For example, the attachment of N-linked glycans to nascent proteins in the endoplasmic reticulum facilitates proper folding, whereas retention of high mannose glycans on misfolded glycoproteins serves as a signal for retrotranslocation and ubiquitin-mediated proteasomal degradation. Here we examine the substrate specificity of the only family of ubiquitin ligase subunits thought to target glycoproteins through their attached glycans. The five proteins comprising this FBA family (FBXO2, FBXO6, FBXO17, FBXO27, and FBXO44) contain a conserved G domain that mediates substrate binding. Using a variety of complementary approaches, including glycan arrays, we show that each family member has differing specificity for glycosylated substrates. Collectively, the F-box proteins in the FBA family bind high mannose and sulfated glycoproteins, with one FBA protein, FBX044, failing to bind any glycans on the tested arrays. Site-directed mutagenesis of two aromatic amino acids in the G domain demonstrated that the hydrophobic pocket created by these amino acids is necessary for high affinity glycan binding. All FBA proteins co-precipitated components of the canonical SCF complex (Skp1, Cullin1, and Rbx1), yet FBXO2 bound very little Cullin1, suggesting that FBXO2 may exist primarily as a heterodimer with Skp1. Using subunit-specific antibodies, we further demonstrate marked divergence in tissue distribution and developmental expression. These differences in substrate recognition, SCF complex formation, and tissue distribution suggest that FBA proteins play diverse roles in glycoprotein quality control.

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Authors
Glenn, K. A., Nelson, R. F., Wen, H. M., Mallinger, A. J., & Paulson, H. L.
Affiliations
  • Veterans Health Administration (US)
  • University of Iowa Hospitals and Clinics (US)
  • Kansas City University (US)
  • Michigan Medicine (US)
Corresponding author
Glenn, Kevin A.
Venue
Journal of Biological Chemistry
Publisher
Elsevier BV
Published
2008-05
Volume
283
Issue
19
Pages
12717-12729
ISSN
0021-9258 (ISSN-L)
OpenAlex
W2007169599
Cited by
86
References
73
Citations by year
2026: 5 · 2025: 5 · 2024: 5 · 2023: 4 · 2022: 1 · 2021: 7 · 2020: 6 · 2019: 4 · 2018: 3 · 2017: 6 · 2016: 4 · 2015: 8 · 2014: 4 · 2013: 3 · 2012: 6
FWCI
1.92
Type
journal article
Language
en
Open access
Locations
  • Journal of Biological Chemistry (publishedVersion, cc-by)
  • Europe PMC (PubMed Central) (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
Funders
  • National Institutes of Health
  • National Institute of General Medical Sciences (T32GM007337)
  • National Institute of Neurological Disorders and Stroke (R01NS047535, F30NS047872, NS047872, R01 NS47535-01)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Ubiquitin and proteasome pathways
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Glycosylation and Glycoproteins Research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Endoplasmic Reticulum Stress and Disease
Keywords
ubiquitin ligase, F-box protein, substrate specificity, glycan binding, high-mannose glycans, sulfated glycoproteins, G-domain, SCF ubiquitin ligase, CUL1, Skp1, RBX1, tissue expression, developmental gene expression, glycoprotein quality control, N-glycosylation, retrotranslocation, proteasomal degradation
MeSH terms
Animals, Cell Line, Chlorocebus aethiops, Multigene Family, Glycoproteins, Humans, Mannose, Models, Biological, Polysaccharides, Protein Binding, Substrate Specificity, Gene Expression Regulation, Enzymologic, Lectins, F-Box Proteins, SKP Cullin F-Box Protein Ligases, Mice, Hydrophobic and Hydrophilic Interactions

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