Structure of proteasome ubiquitin receptor hRpn13 and its activation by the scaffolding protein hRpn2.
Rpn13 is a subunit of the proteasome that serves as a receptor for both ubiquitin and Uch37, one of the proteasome's three deubiquitinating enzymes. We have determined the structure of full-length human Rpn13 (hRpn13). Unexpectedly, Rpn13's ubiquitin- and Uch37-binding domains pack against each other when it is not incorporated into the proteasome. This intramolecular interaction reduces hRpn13's affinity for ubiquitin. We find that hRpn13 binding to the proteasome scaffolding protein hRpn2/S1 abrogates its interdomain interactions, thus activating hRpn13 for ubiquitin binding. hRpn13's Uch37-binding domain, a previously unknown fold, contains nine alpha helices. We have mapped its Uch37-binding surface to a region rich in charged amino acids. Altogether, our results provide mechanistic insights into hRpn13's functional activities with Uch37 and ubiquitin and suggest that its role as a ubiquitin receptor is finely tuned for proteasome targeting.
Chen
Xiang
X
Department of Biochemistry, University of Minnesota, Minneapolis, MN 55455, USA.
Lee
Byung-Hoon
BH
Finley
Daniel
D
Walters
Kylie J
KJ
D000595:Amino Acid Sequence; D001665:Binding Sites; D002268:Carboxypeptidases / Q000378:metabolism; D006602:Hexosyltransferases; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D008562:Membrane Glycoproteins / Q000737:chemistry* / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D046988:Proteasome Endopeptidase Complex / Q000737:chemistry* / Q000378:metabolism; D011487:Protein Conformation; D011499:Protein Processing, Post-Translational; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D013329:Structure-Activity Relationship; D025801:Ubiquitin / Q000378:metabolism; D043222:Ubiquitin Thiolesterase; D054875:Ubiquitination
C500781:ADRM1 protein, human; D047908:Intracellular Signaling Peptides and Proteins; D008562:Membrane Glycoproteins; D011993:Recombinant Fusion Proteins; D025801:Ubiquitin; D006602:Hexosyltransferases; C517590:RPN2 protein, human; D002268:Carboxypeptidases; C421245:UCHL5 protein, human; D043222:Ubiquitin Thiolesterase; D046988:Proteasome Endopeptidase Complex
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