title: Structure of proteasome ubiquitin receptor hRpn13 and its activation by the scaffolding protein hRpn2.
abstract: 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.
authors.affiliation: Department of Biochemistry, University of Minnesota, Minneapolis, MN 55455, USA.
authors.forename: Xiang
authors.initials: X
authors.lastname: Chen
authors.forename: Byung-Hoon
authors.initials: BH
authors.lastname: Lee
authors.forename: Daniel
authors.initials: D
authors.lastname: Finley
authors.forename: Kylie J
authors.initials: KJ
authors.lastname: Walters
mesh_terms: 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
chemical_list: 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
references.citation: Nature. 1996 Dec 19-26;384(6610):638-41
references.pmid: 8967952
references.citation: J Biol Chem. 2001 Jul 27;276(30):27936-43
references.pmid: 11369780
references.citation: Biochem Biophys Res Commun. 2009 Dec 18;390(3):855-60
references.pmid: 19836345
references.citation: Anal Biochem. 1979 Apr 1;94(1):75-81
references.pmid: 37780
references.citation: Mol Cell. 2009 Aug 14;35(3):280-90
references.pmid: 19683493
references.citation: J Am Chem Soc. 2001 Oct 10;123(40):9843-7
references.pmid: 11583547
references.citation: Cell. 2009 Apr 3;137(1):133-45
references.pmid: 19345192
references.citation: Nature. 2006 Nov 16;444(7117):383-6
references.pmid: 17051159
references.citation: Nature. 2007 Oct 25;449(7165):1078-82
references.pmid: 17960247
references.citation: J Magn Reson. 2003 Jan;160(1):65-73
references.pmid: 12565051
references.citation: Annu Rev Biochem. 2009;78:477-513
references.pmid: 19489727
references.citation: J Magn Reson. 2007 Feb;184(2):185-95
references.pmid: 17084097
references.citation: J Biomol NMR. 1999 Mar;13(3):289-302
references.pmid: 10212987
references.citation: EMBO J. 2006 Oct 4;25(19):4524-36
references.pmid: 16990800
references.citation: Science. 2005 Feb 25;307(5713):1317-21
references.pmid: 15731457
references.citation: J Mol Biol. 2007 Mar 16;367(1):204-11
references.pmid: 17240395
references.citation: J Am Chem Soc. 2006 Apr 5;128(13):4389-97
references.pmid: 16569016
references.citation: Science. 2005 Jul 1;309(5731):127-30
references.pmid: 15994556
references.citation: Methods Enzymol. 2005;399:21-36
references.pmid: 16338346
references.citation: Bioinformatics. 2008 Dec 1;24(23):2780-1
references.pmid: 18818215
references.citation: Nat Genet. 2006 Mar;38(3):285-93
references.pmid: 16501559
references.citation: Nat Cell Biol. 2006 Jul;8(7):700-10
references.pmid: 16799550
references.citation: J Biomol NMR. 1995 Nov;6(3):277-93
references.pmid: 8520220
references.citation: Nat Cell Biol. 2006 Sep;8(9):994-1002
references.pmid: 16906146
references.citation: Nature. 2007 Dec 13;450(7172):983-90
references.pmid: 18075577
references.citation: J Cell Biol. 2007 May 21;177(4):613-24
references.pmid: 17502423
references.citation: J Mol Biol. 2002 Dec 6;324(4):637-47
references.pmid: 12460567
references.citation: Nat Struct Mol Biol. 2009 Aug;16(8):883-9
references.pmid: 19620964
references.citation: Nat Cell Biol. 2007 Apr;9(4):422-7
references.pmid: 17310239
references.citation: Nature. 2008 May 22;453(7194):548-52
references.pmid: 18497827
references.citation: J Biol Chem. 2004 Oct 1;279(40):41414-21
references.pmid: 15254040
references.citation: J Mol Biol. 2005 May 6;348(3):727-39
references.pmid: 15826667
references.citation: Biochemistry. 2000 May 9;39(18):5355-65
references.pmid: 10820006
references.citation: J Biomol NMR. 1995 Jul;6(1):1-10
references.pmid: 22911575
references.citation: Biochemistry. 2007 Mar 20;46(11):3553-65
references.pmid: 17323924
references.citation: J Biol Chem. 2007 Jun 15;282(24):17375-86
references.pmid: 17426036
references.citation: J Biol Chem. 1994 Mar 11;269(10):7059-61
references.pmid: 8125911
references.citation: EMBO J. 1998 Oct 15;17(20):5964-73
references.pmid: 9774340
references.citation: Nature. 2008 May 22;453(7194):481-8
references.pmid: 18497817
references.citation: EMBO J. 2006 Oct 18;25(20):4877-87
references.pmid: 17006537
references.citation: J Mol Biol. 1994 Feb 18;236(2):601-9
references.pmid: 8107144
references.citation: Nature. 2006 Apr 27;440(7088):1227-30
references.pmid: 16642002
references.citation: EMBO J. 2009 Feb 18;28(4):359-71
references.pmid: 19153599
references.citation: EMBO J. 2006 Dec 13;25(24):5742-53
references.pmid: 17139257
references.citation: Methods Enzymol. 1996;266:554-71
references.pmid: 8743706
references.citation: Nature. 2007 Dec 6;450(7171):838-44
references.pmid: 18026086
references.citation: J Biol Chem. 1997 Nov 7;272(45):28438-46
references.pmid: 9353303
references.citation: Proc Natl Acad Sci U S A. 2001 Apr 10;98(8):4569-74
references.pmid: 11283351