Characterizing polyubiquitinated forms of the neurodegenerative ubiquitin mutant UBB+1

FEBS Lett. 2016 Dec;590(24):4573-4585. doi: 10.1002/1873-3468.12484. Epub 2016 Nov 22.

Abstract

The ubiquitin mutant UBB+1 has been identified as a hallmark of neurodegenerative diseases. In this study, we characterize polyubiquitinated forms of UBB+1 in vitro and from patient samples. The ability of UBB+1 to be readily ubiquitinated by several E2 enzymes provided a mechanism for the controlled synthesis and purification of defined conjugates. This allowed us to utilize polyUb-UBB+1 conjugates for biochemical assays, as well as solution NMR. Coupled with our immunoassay for detection of ubiquitinated forms of UBB+1 in patient blood samples, we gain a clearer picture of the molecular mechanisms underlying neurodegenerative diseases.

Keywords: UBB +1; biomolecular NMR; deubiquitinase; neurodegeneration; polyubiquitin.

Publication types

  • Letter

MeSH terms

  • Aged
  • Alzheimer Disease / genetics*
  • Alzheimer Disease / metabolism
  • Alzheimer Disease / pathology
  • Case-Control Studies
  • Cysteine Endopeptidases / chemistry
  • Cysteine Endopeptidases / genetics
  • Cysteine Endopeptidases / metabolism
  • Deubiquitinating Enzymes
  • Endopeptidases / chemistry
  • Endopeptidases / genetics
  • Endopeptidases / metabolism
  • Endosomal Sorting Complexes Required for Transport / chemistry
  • Endosomal Sorting Complexes Required for Transport / genetics
  • Endosomal Sorting Complexes Required for Transport / metabolism
  • Female
  • Fluoresceins / chemistry
  • Fluorescent Dyes / chemistry
  • Gene Expression Regulation
  • Humans
  • Male
  • Middle Aged
  • Models, Molecular
  • Neurons / metabolism*
  • Neurons / pathology
  • Nuclear Magnetic Resonance, Biomolecular
  • Polyubiquitin / chemistry*
  • Polyubiquitin / genetics
  • Polyubiquitin / metabolism
  • Proteasome Endopeptidase Complex / metabolism
  • Protein Isoforms / chemistry
  • Protein Isoforms / genetics
  • Protein Isoforms / metabolism
  • Protein Structure, Secondary
  • Staining and Labeling
  • Ubiquitin / chemistry*
  • Ubiquitin / genetics
  • Ubiquitin / metabolism
  • Ubiquitin Thiolesterase / chemistry
  • Ubiquitin Thiolesterase / genetics
  • Ubiquitin Thiolesterase / metabolism
  • Ubiquitin-Specific Proteases / chemistry
  • Ubiquitin-Specific Proteases / genetics
  • Ubiquitin-Specific Proteases / metabolism
  • Ubiquitination

Substances

  • Endosomal Sorting Complexes Required for Transport
  • Fluoresceins
  • Fluorescent Dyes
  • Protein Isoforms
  • STAMBP protein, human
  • UBB protein, human
  • Ubiquitin
  • Polyubiquitin
  • fluorescein 5-maleimide
  • Endopeptidases
  • Deubiquitinating Enzymes
  • OTUB1 protein, human
  • OTUD3 protein, human
  • USP2 protein, human
  • Ubiquitin Thiolesterase
  • Ubiquitin-Specific Proteases
  • Cysteine Endopeptidases
  • Proteasome Endopeptidase Complex