Q9HAU4 (SMUF2_HUMAN) Reviewed, UniProtKB/Swiss-Prot

Names and origin

Protein names

Recommended name:
E3 ubiquitin-protein ligase SMURF2
Short name=hSMURF2
EC=6.3.2.-

Alternative name(s):
SMAD ubiquitination regulatory factor 2
SMAD-specific E3 ubiquitin-protein ligase 2

Gene names

Name:

SMURF2

Organism

Homo sapiens (Human) [Reference proteome]

Taxonomic identifier

9606 [NCBI]

Taxonomic lineage

cellular organisms › Eukaryota › Opisthokonta › Metazoa › Eumetazoa › Bilateria › Deuterostomia › Chordata › Craniata › Vertebrata › Gnathostomata › Teleostomi › Euteleostomi › Sarcopterygii › Tetrapoda › Amniota › Mammalia › Theria › Eutheria › Euarchontoglires › Primates › Haplorrhini › Simiiformes › Catarrhini › Hominoidea › Hominidae › Homininae › Homo

Protein attributes

Sequence length	748 AA.
Sequence status	Complete.
Protein existence	Evidence at protein level

General annotation (Comments)

Function	E3 ubiquitin-protein ligase which accepts ubiquitin from an E2 ubiquitin-conjugating enzyme in the form of a thioester and then directly transfers the ubiquitin to targeted substrates. Interacts with SMAD1 and SMAD7 in order to trigger their ubiquitination and proteasome-dependent degradation. In addition, interaction with SMAD7 activates autocatalytic degradation, which is prevented by interaction with SCYE1. Forms a stable complex with the TGF-beta receptor-mediated phosphorylated SMAD2 and SMAD3. In this way, SMAD2 may recruit substrates, such as SNON, for ubiquitin-mediated degradation. Enhances the inhibitory activity of SMAD7 and reduces the transcriptional activity of SMAD2. Coexpression of SMURF2 with SMAD1 results in considerable decrease in steady-state level of SMAD1 protein and a smaller decrease of SMAD2 level. Ref.5 Ref.6
Enzyme regulation	Activated by NDFIP1- and NDFIP2-binding.
Pathway	Protein modification; protein ubiquitination.
Subunit structure	Interacts (via WW domains) with SMAD1. Interacts (via WW domains) with SMAD2 (via PY-motif). Interacts (via WW domains) with SMAD3 (via PY-motif). Interacts with SMAD6. Interacts with SMAD7 (via PY-motif) and TGFBR1; SMAD7 recruits SMURF2 to the TGF-beta receptor and regulates its degradation. Does not interact with SMAD4; SMAD4 lacks a PY-motif. Interacts with AIMP1. Interacts with STAMBP and RNF11. Interacts with NDFIP1 and NDFIP2 Probable; this interaction activates the E3 ubiquitin-protein ligase. Ref.1 Ref.5 Ref.8 Ref.9 Ref.10 Ref.13
Subcellular location	Nucleus. Cytoplasm. Cell membrane. Membrane raft. Note: Cytoplasmic in the presence of SMAD7. Co-localizes with CAV1, SMAD7 and TGF-beta receptor in membrane rafts. Ref.6
Tissue specificity	Widely expressed.
Domain	The second and third WW domains are responsible for interaction with the PY-motif of R-SMAD (SMAD1, SMAD2 and SMAD3). Ref.15 The C2 domain is involved in autoinhibition of the catalytic activity by interacting with the HECT domain. Ref.15
Post-translational modification	Auto-ubiquitinated and ubiquitinated in the presence of RNF11 and UBE2D1. Ubiquitinated by the SCF(FBXL15) complex, leading to its degradation by the proteasome. Ref.11 Ref.12
Sequence similarities	Contains 1 C2 domain. Contains 1 HECT (E6AP-type E3 ubiquitin-protein ligase) domain. Contains 3 WW domains.

Ontologies

Keywords
Biological process	Ubl conjugation pathway
Cellular component	Cell membrane Cytoplasm Membrane Nucleus
Domain	Repeat
Molecular function	Ligase
PTM	Ubl conjugation
Technical term	3D-structure Complete proteome Reference proteome
Gene Ontology (GO)
Biological_process	BMP signaling pathway Traceable author statement. Source: Reactome negative regulation of transcription from RNA polymerase II promoter Traceable author statement. Source: Reactome negative regulation of transforming growth factor beta receptor signaling pathway Traceable author statement. Source: Reactome protein ubiquitination involved in ubiquitin-dependent protein catabolic process Inferred from Biological aspect of Ancestor. Source: RefGenome transcription initiation from RNA polymerase II promoter Traceable author statement. Source: Reactome transforming growth factor beta receptor signaling pathway Inferred from Biological aspect of Ancestor. Source: RefGenome ubiquitin-dependent SMAD protein catabolic process Inferred from direct assay PubMed 19122240. Source: BHF-UCL
Cellular_component	cytosol Traceable author statement. Source: Reactome membrane raft Inferred from electronic annotation. Source: UniProtKB-SubCell nucleoplasm Traceable author statement. Source: Reactome plasma membrane Inferred from electronic annotation. Source: UniProtKB-SubCell ubiquitin ligase complex Inferred from direct assay Ref.9. Source: UniProtKB
Molecular_function	ubiquitin-protein ligase activity Inferred from direct assay Ref.9. Source: UniProtKB
Complete GO annotation...

Binary interactions

With	Entry	#Exp.	IntAct	Notes
itself		5	EBI-396727,EBI-396727
RNF11	Q9Y3C5	3	EBI-396727,EBI-396669
UBB	P0CG47	4	EBI-396727,EBI-413034

Sequence annotation (Features)

Feature key

Position(s)

Length

Description

Graphical view

Feature identifier

Molecule processing

Chain

1 – 748

748

E3 ubiquitin-protein ligase SMURF2

PRO_0000120329

Regions

Domain

1 – 98

98

C2

Domain

157 – 190

34

WW 1

Domain

251 – 284

34

WW 2

Domain

297 – 330

34

WW 3

Domain

414 – 748

335

HECT

Sites

Active site

716

1

Glycyl thioester intermediate By similarity

Experimental info

Mutagenesis

29

1

F → A: Increases autoubiquitination; when associated with A-30. Ref.15

Mutagenesis

30

1

F → A: Increases autoubiquitination; when associated with A-29. Ref.15

Mutagenesis

56

1

T → A: Increases autoubiquitination; when associated with A-57. Ref.15

Mutagenesis

57

1

L → A: Increases autoubiquitination; when associated with A-56. Ref.15

Mutagenesis

251 – 284

34

Missing: Abolishes interaction with SMAD2 and SMAD7. Ref.1 Ref.2

Mutagenesis

297 – 330

34

Missing: Abolishes interaction with SMAD7. Ref.1

Mutagenesis

535

1

W → A: Loss of catalytic activity. Ref.13

Mutagenesis

535

1

W → D: Loss of catalytic activity. Ref.13

Mutagenesis

547

1

H → A: Partial loss of catalytic activity. Ref.13

Mutagenesis

547

1

H → F or I: Activates autocatalytic activity. Ref.13

Mutagenesis

581

1

Y → A: Loss of catalytic activity. Ref.13

Mutagenesis

716

1

C → A: Increases Smad7-bound TGF-beta receptors in membrane rafts. Ref.2 Ref.3 Ref.6

Mutagenesis

716

1

C → G: Loss of activity. Loss of ability to ubiquitinate SMAD1 and SMAD2 and no down-regulation of SMAD1 and SMAD2 protein levels. Ref.2 Ref.3 Ref.6

Sequence conflict

6

1

G → R in AAG45422. Ref.2

Secondary structure

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748

Helix Strand Turn

Details...

Beta strand

12 – 23

12

Beta strand

35 – 41

7

Turn

42 – 44

3

Beta strand

48 – 50

3

Beta strand

60 – 69

10

Beta strand

75 – 81

7

Helix

82 – 87

6

Beta strand

93 – 99

7

Helix

101 – 110

10

Beta strand

113 – 115

3

Beta strand

131 – 138

8

Beta strand

258 – 262

5

Turn

263 – 265

3

Beta strand

266 – 271

6

Turn

272 – 275

4

Beta strand

276 – 280

5

Beta strand

282 – 284

3

Beta strand

286 – 288

3

Helix

293 – 295

3

Beta strand

301 – 307

7

Beta strand

309 – 311

3

Beta strand

313 – 317

5

Turn

318 – 321

4

Beta strand

322 – 326

5

Turn

328 – 330

3

Helix

372 – 386

15

Beta strand

392 – 397

6

Helix

402 – 411

10

Helix

415 – 419

5

Beta strand

420 – 426

7

Helix

434 – 449

16

Helix

452 – 454

3

Beta strand

455 – 460

6

Beta strand

463 – 469

7

Helix

473 – 475

3

Helix

479 – 495

17

Helix

506 – 512

7

Helix

522 – 525

4

Helix

527 – 538

12

Turn

542 – 544

3

Beta strand

549 – 555

7

Beta strand

558 – 565

8

Turn

574 – 577

4

Helix

578 – 590

13

Turn

591 – 594

4

Helix

595 – 608

14

Helix

611 – 614

4

Helix

619 – 627

9

Beta strand

629 – 631

3

Helix

634 – 639

6

Beta strand

641 – 646

6

Helix

651 – 662

12

Helix

665 – 676

12

Beta strand

679 – 681

3

Helix

686 – 688

3

Beta strand

698 – 702

5

Beta strand

712 – 714

3

Helix

715 – 717

3

Beta strand

719 – 722

4

Helix

728 – 739

12

Sequences

	Sequence		Length	Mass (Da)
	Q9HAU4 [UniParc]. Last modified March 1, 2001. Version 1. Checksum: 3042B443A3755762	FASTA	748	86,196
	10 20 30 40 50 60 MSNPGGRRNG PVKLRLTVLC AKNLVKKDFF RLPDPFAKVV VDGSGQCHST DTVKNTLDPK 70 80 90 100 110 120 WNQHYDLYIG KSDSVTISVW NHKKIHKKQG AGFLGCVRLL SNAINRLKDT GYQRLDLCKL 130 140 150 160 170 180 GPNDNDTVRG QIVVSLQSRD RIGTGGQVVD CSRLFDNDLP DGWEERRTAS GRIQYLNHIT 190 200 210 220 230 240 RTTQWERPTR PASEYSSPGR PLSCFVDENT PISGTNGATC GQSSDPRLAE RRVRSQRHRN 250 260 270 280 290 300 YMSRTHLHTP PDLPEGYEQR TTQQGQVYFL HTQTGVSTWH DPRVPRDLSN INCEELGPLP 310 320 330 340 350 360 PGWEIRNTAT GRVYFVDHNN RTTQFTDPRL SANLHLVLNR QNQLKDQQQQ QVVSLCPDDT 370 380 390 400 410 420 ECLTVPRYKR DLVQKLKILR QELSQQQPQA GHCRIEVSRE EIFEESYRQV MKMRPKDLWK 430 440 450 460 470 480 RLMIKFRGEE GLDYGGVARE WLYLLSHEML NPYYGLFQYS RDDIYTLQIN PDSAVNPEHL 490 500 510 520 530 540 SYFHFVGRIM GMAVFHGHYI DGGFTLPFYK QLLGKSITLD DMELVDPDLH NSLVWILEND 550 560 570 580 590 600 ITGVLDHTFC VEHNAYGEII QHELKPNGKS IPVNEENKKE YVRLYVNWRF LRGIEAQFLA 610 620 630 640 650 660 LQKGFNEVIP QHLLKTFDEK ELELIICGLG KIDVNDWKVN TRLKHCTPDS NIVKWFWKAV 670 680 690 700 710 720 EFFDEERRAR LLQFVTGSSR VPLQGFKALQ GAAGPRLFTI HQIDACTNNL PKAHTCFNRI 730 740 DIPPYESYEK LYEKLLTAIE ETCGFAVE

References

	« Hide 'large scale' referencesShow 'large scale' references »
[1]	"Smad7 binds to Smurf2 to form an E3 ubiquitin ligase that targets the TGF-beta receptor for degradation." Kavsak P., Rasmussen R.K., Causing C.G., Bonni S., Zhu H., Thomsen G.H., Wrana J.L. Mol. Cell 6:1365-1375(2000) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [MRNA], MUTAGENESIS OF 251-PRO--VAL-284 AND 297-GLY--LEU-330, INTERACTION WITH SMAD7 AND TGFBR1.
[2]	"Smurf2 Is a ubiquitin E3 ligase mediating proteasome-dependent degradation of Smad2 in transforming growth factor-beta signaling." Lin X., Liang M., Feng X.-H. J. Biol. Chem. 275:36818-36822(2000) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [MRNA], MUTAGENESIS OF 251-PRO--VAL-284 AND CYS-716.
[3]	"Regulation of Smad degradation and activity by Smurf2, an E3 ubiquitin ligase." Zhang Y., Chang C., Gehling D.J., Hemmati-Brivanlou A., Derynck R. Proc. Natl. Acad. Sci. U.S.A. 98:974-979(2001) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [MRNA], MUTAGENESIS OF CYS-716.
[4]	"The status, quality, and expansion of the NIH full-length cDNA project: the Mammalian Gene Collection (MGC)." The MGC Project Team Genome Res. 14:2121-2127(2004) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE MRNA]. Tissue: Brain.
[5]	"TGF-beta induces assembly of a Smad2-Smurf2 ubiquitin ligase complex that targets SnoN for degradation." Bonni S., Wang H.R., Causing C.G., Kavsak P., Stroschein S.L., Luo K., Wrana J.L. Nat. Cell Biol. 3:587-595(2001) [PubMed] [Europe PMC] [Abstract] Cited for: FUNCTION, INTERACTION WITH SMAD2; SMAD3 AND SNON.
[6]	"Distinct endocytic pathways regulate TGF-beta receptor signalling and turnover." Di Guglielmo G.M., Le Roy C., Goodfellow A.F., Wrana J.L. Nat. Cell Biol. 5:410-421(2003) [PubMed] [Europe PMC] [Abstract] Cited for: FUNCTION, SUBCELLULAR LOCATION, MUTAGENESIS OF CYS-716.
[7]	Erratum Di Guglielmo G.M., Le Roy C., Goodfellow A.F., Wrana J.L. Nat. Cell Biol. 5:680-680(2003)
[8]	"The RING-H2 protein RNF11 is overexpressed in breast cancer and is a target of Smurf2 E3 ligase." Subramaniam V., Li H., Wong M.J., Kitching R., Attisano L., Wrana J., Zubovits J., Burger A.M., Seth A.K. Br. J. Cancer 89:1538-1544(2003) [PubMed] [Europe PMC] [Abstract] Cited for: INTERACTION WITH RNF11.
[9]	"An RNF11: Smurf2 complex mediates ubiquitination of the AMSH protein." Li H., Seth A.K. Oncogene 23:1801-1808(2004) [PubMed] [Europe PMC] [Abstract] Cited for: INTERACTION WITH STAMBP AND RNF11.
[10]	"AIMP1/p43 downregulates TGF-beta signaling via stabilization of smurf2." Lee Y.S., Han J.M., Son S.H., Choi J.W., Jeon E.J., Bae S.-C., Park Y.I., Kim S. Biochem. Biophys. Res. Commun. 371:395-400(2008) [PubMed] [Europe PMC] [Abstract] Cited for: INTERACTION WITH AIMP1.
[11]	"Control of the activity of WW-HECT domain E3 ubiquitin ligases by NDFIP proteins." Mund T., Pelham H.R. EMBO Rep. 10:501-507(2009) [PubMed] [Europe PMC] [Abstract] Cited for: ACTIVATION BY NDFIP1 AND NDFIP2, AUTOUBIQUITINATION.
[12]	"SCF(FBXL15) regulates BMP signalling by directing the degradation of HECT-type ubiquitin ligase Smurf1." Cui Y., He S., Xing C., Lu K., Wang J., Xing G., Meng A., Jia S., He F., Zhang L. EMBO J. 30:2675-2689(2011) [PubMed] [Europe PMC] [Abstract] Cited for: UBIQUITINATION.
[13]	"Regulation of Smurf2 ubiquitin ligase activity by anchoring the E2 to the HECT domain." Ogunjimi A.A., Briant D.J., Pece-Barbara N., Le Roy C., Di Guglielmo G.M., Kavsak P., Rasmussen R.K., Seet B.T., Sicheri F., Wrana J.L. Mol. Cell 19:297-308(2005) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.1 ANGSTROMS) OF 369-748, INTERACTION WITH SMAD7, MUTAGENESIS OF TRP-535; HIS-547 AND TYR-581.
[14]	"An expanded WW domain recognition motif revealed by the interaction between Smad7 and the E3 ubiquitin ligase Smurf2." Chong P.A., Lin H., Wrana J.L., Forman-Kay J.D. J. Biol. Chem. 281:17069-17075(2006) [PubMed] [Europe PMC] [Abstract] Cited for: STRUCTURE BY NMR OF 297-333 IN COMPLEX WITH SMAD7.
[15]	"Autoinhibition of the HECT-type ubiquitin ligase Smurf2 through its C2 domain." Wiesner S., Ogunjimi A.A., Wang H.R., Rotin D., Sicheri F., Wrana J.L., Forman-Kay J.D. Cell 130:651-662(2007) [PubMed] [Europe PMC] [Abstract] Cited for: STRUCTURE BY NMR OF 10-140, AUTOINHIBITION BY C2 DOMAIN, MUTAGENESIS OF PHE-29; PHE-30; THR-56 AND LEU-57.
+	Additional computationally mapped references.

Cross-references

Sequence databases

EMBL
GenBank
DDBJ

AF310676 mRNA. Translation: AAG45422.1.
AF301463 mRNA. Translation: AAG25641.1.
AY014180 mRNA. Translation: AAG50421.1.
BC093876 mRNA. Translation: AAH93876.1.
BC111945 mRNA. Translation: AAI11946.1.

IPI

IPI00329664.

RefSeq

NP_073576.1. NM_022739.3.

UniGene

Hs.741194.

3D structure databases

PDBe
RCSB PDB
PDBj

Entry	Method	Resolution (Å)	Chain	Positions	PDBsum
1ZVD	X-ray	2.10	A	369-748	[»]
2DJY	NMR	-	A	297-333	[»]
2JQZ	NMR	-	A	10-140	[»]
2KXQ	NMR	-	A	250-333	[»]
2LTZ	NMR	-	A	297-333	[»]

ProteinModelPortal

Q9HAU4.

ModBase

Search...

Protein-protein interaction databases

DIP

DIP-33061N.

IntAct

Q9HAU4. 7 interactions.

MINT

MINT-1180022.

STRING

9606.ENSP00000262435.

PTM databases

PhosphoSite

Q9HAU4.

Polymorphism databases

DMDM

17865624.

Proteomic databases

PaxDb

Q9HAU4.

PRIDE

Q9HAU4.

Protocols and materials databases

StructuralBiologyKnowledgebase

Search...

Genome annotation databases

Ensembl

ENST00000262435; ENSP00000262435; ENSG00000108854.
ENST00000573633; ENSP00000461327; ENSG00000262909.

GeneID

64750.

KEGG

hsa:64750.

UCSC

uc002jep.1. human.

Organism-specific databases

CTD

64750.

GeneCards

GC17M062540.

HGNC

HGNC:16809. SMURF2.

MIM

605532. gene.

neXtProt

NX_Q9HAU4.

PharmGKB

PA134985524.

GenAtlas

Search...

Phylogenomic databases

eggNOG

COG5021.

HOGENOM

HOG000208451.

HOVERGEN

HBG004134.

InParanoid

Q9HAU4.

KO

K04678.

OMA

ANQAARP.

OrthoDB

EOG47PX58.

PhylomeDB

Q9HAU4.

Enzyme and pathway databases

Pathway_Interaction_DB

bmppathway. BMP receptor signaling.
tgfbrpathway. TGF-beta receptor signaling.

Reactome

REACT_111102. Signal Transduction.
REACT_6900. Immune System.
REACT_71. Gene Expression.

UniPathway

UPA00143.

Gene expression databases

ArrayExpress

Q9HAU4.

Bgee

Q9HAU4.

CleanEx

HS_SMURF2.

Genevestigator

Q9HAU4.

GermOnline

ENSG00000108854. Homo sapiens.

Family and domain databases

InterPro

IPR000008. C2_Ca-dep.
IPR008973. C2_Ca/lipid-bd_dom_CaLB.
IPR018029. C2_membr_targeting.
IPR024928. E3_ub_ligase_SMURF1.
IPR000569. HECT.
IPR001202. WW_dom.
[Graphical view]

Pfam

PF00168. C2. 1 hit.
PF00632. HECT. 1 hit.
PF00397. WW. 3 hits.
[Graphical view]

PIRSF

PIRSF001569. E3_ub_ligase_SMURF1. 1 hit.

SMART

SM00239. C2. 1 hit.
SM00119. HECTc. 1 hit.
SM00456. WW. 3 hits.
[Graphical view]

SUPFAM

SSF49562. C2_CaLB. 1 hit.
SSF56204. HECT. 1 hit.
SSF51045. WW_Rsp5_WWP. 3 hits.

PROSITE

PS50004. C2. 1 hit.
PS50237. HECT. 1 hit.
PS01159. WW_DOMAIN_1. 1 hit.
PS50020. WW_DOMAIN_2. 3 hits.
[Graphical view]

ProtoNet

Search...

Other

ChiTaRS

SMURF2. human.

EvolutionaryTrace

Q9HAU4.

GenomeRNAi

64750.

NextBio

66708.

SOURCE

Search...

Entry information

Entry name

SMUF2_HUMAN

Accession

Primary (citable) accession number: Q9HAU4
Secondary accession number(s): Q52LL1, Q9H260

Entry history

Integrated into UniProtKB/Swiss-Prot:	December 13, 2001
Last sequence update:	March 1, 2001
Last modified:	May 1, 2013
This is version 128 of the entry and version 1 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)

Annotation program

Chordata Protein Annotation Program

Disclaimer

Any medical or genetic information present in this entry is provided for research, educational and informational purposes only. It is not in any way intended to be used as a substitute for professional medical advice, diagnosis, treatment or care.