NEDP1, a highly conserved cysteine protease that deNEDDylates Cullins.
The ubiquitin-like protein NEDD8 is essential for activity of SCF-like ubiquitin ligase complexes. Here we identify and characterize NEDP1, a human NEDD8-specific protease. NEDP1 is highly conserved throughout evolution and equivalent proteins are present in yeast, plants, insects, and mammals. Bacterially expressed NEDP1 is capable of processing NEDD8 in vitro to expose the diglycine motif required for conjugation and can deconjugate NEDD8 from modified substrates. NEDP1 appears to be specific for NEDD8 as neither ubiquitin nor SUMO bearing COOH-terminal extensions are utilized as substrates. Inhibition studies and mutagenesis indicate that NEDP1 is a cysteine protease with sequence similarities to SUMO-specific proteases and the class of viral proteases typified by the adenovirus protease. In vivo NEDP1 deconjugates NEDD8 from a wide variety of substrates including the cullin component of SCF-like complexes. Thus NEDP1 is likely to play an important role in ubiquitin-mediated proteolysis by controlling the activity of SCF complexes.
Centre for Biomolecular Sciences, School of Biology, University of St. Andrews, North Haugh, St. Andrews, Fife KY169AL, United Kingdom.
Heidi M
HM
Mendoza
Lin-Nan
LN
Shen
Catherine
C
Botting
Alan
A
Lewis
Jingwen
J
Chen
Barbara
B
Ink
Ronald T
RT
Hay
D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D019556:COS Cells; D018797:Cell Cycle Proteins / Q000378:metabolism*; D003001:Cloning, Molecular; D044842:Cullin Proteins; D003546:Cysteine Endopeptidases / Q000737:chemistry* / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D010450:Endopeptidases; D005822:Genetic Vectors; D005982:Glutathione Transferase / Q000378:metabolism; D006801:Humans; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D000075687:NEDD8 Protein; D010957:Plasmids / Q000378:metabolism; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity; D014018:Tissue Distribution; D014162:Transfection; D025801:Ubiquitin / Q000378:metabolism; D014452:Ubiquitins / Q000378:metabolism*
D018797:Cell Cycle Proteins; C100318:Cullin 1; D044842:Cullin Proteins; D000075687:NEDD8 Protein; C082860:NEDD8 protein, human; D011993:Recombinant Fusion Proteins; D025801:Ubiquitin; D014452:Ubiquitins; D005982:Glutathione Transferase; D010450:Endopeptidases; D003546:Cysteine Endopeptidases; C477008:SENP8 protein, human