Reviewed, UniProtKB/Swiss-Prot O88339 (EPN1_RAT)

Names and origin

Protein names

Recommended name:
    Epsin-1
Alternative name(s):
    EPS-15-interacting protein 1

Gene names

Name:

Epn1

Organism

Rattus norvegicus (Rat)

Taxonomic identifier

10116 [NCBI]

Taxonomic lineage

Eukaryota › Metazoa › Chordata › Craniata › Vertebrata › Euteleostomi › Mammalia › Eutheria › Euarchontoglires › Glires › Rodentia › Sciurognathi › Muroidea › Muridae › Murinae › Rattus

Protein attributes

Sequence length	575 AA.
Sequence status	Complete.
Protein existence	Evidence at protein level.

General annotation (Comments)

Function	Binds to membranes enriched in phosphatidylinositol-4,5-biphosphate (PtdIns(4,5)P2). Modifies membrane curvature and facilitates the formation of clathrin-coated invaginations. Regulates receptor-mediated endocytosis. Ref.1 Ref.7 Ref.10
Subunit structure	Monomer. Binds REPS2 and ITSN1 By similarity. Binds EPS15, clathrin, ZBTB16/ZNF145, AP2A1 and AP2A2. Binds ubiquitinated proteins. Interacts with RALBP1 in a complex also containing NUMB and TFAP2A during interphase and mitosis. Interacts with AP2B1. Ref.1 Ref.7 Ref.2 Ref.3 Ref.4 Ref.5 Ref.6 Ref.9
Subcellular location	Cytoplasm. Cell membrane; Peripheral membrane protein. Nucleus. Membrane › clathrin-coated pit. Note: Associated with the cytoplasmic membrane at sites where clathrin-coated pits are forming. Colocalizes with clathrin and AP-2 in a punctate pattern on the plasma membrane. Colocalizes with clathrin at the Golgi complex. Detected in presynaptic nerve terminals and in synaptosomes. May shuttle to the nucleus when associated with ZBTB16/ZNF145. Ref.1 Ref.7 Ref.6 Ref.9
Tissue specificity	Ubiquitous. Ref.1
Domain	The NPF repeat domain is involved in EPS15 binding. The DPW repeat domain is involved in AP2A2 and clathrin binding. The [DE]-X(1,2)-F-X-X-[FL]-X-X-X-R motif mediates interaction the AP-2 complex subunit AP2B1 By similarity.
Post-translational modification	Ubiquitinated. Ref.8 Phosphorylated on serine and/or threonine residues in mitotic cells. Phosphorylation reduces interaction with REPS2, AP-2 and the membrane fraction. Depolarization of synaptosomes results in dephosphorylation. Ref.3
Sequence similarities	Belongs to the epsin family. Contains 1 ENTH (epsin N-terminal homology) domain. Contains 3 UIM (ubiquitin-interacting motif) repeats.

Ontologies

Keywords
Biological process	Endocytosis
Cellular component	Cell membrane Coated pit Cytoplasm Membrane Nucleus
Domain	Repeat
Ligand	Lipid-binding
PTM	Phosphoprotein Ubl conjugation
Technical term	3D-structure
Gene Ontology (GO)
Biological process	endocytosis Ref.10 Inferred from direct assay. Source: RGD
Cellular component	coated pit Inferred from electronic annotation. Source: UniProtKB-SubCell cytoplasm Inferred from electronic annotation. Source: UniProtKB-SubCell extrinsic to membrane Inferred from electronic annotation. Source: UniProtKB-SubCell nucleus Inferred from direct assay. Source: RGD plasma membrane Inferred from electronic annotation. Source: UniProtKB-KW
Molecular function	lipid binding Inferred from electronic annotation. Source: UniProtKB-KW transcription factor binding Inferred from physical interaction. Source: RGD
Complete GO annotation...

Sequence annotation (Features)

Feature key

Position(s)

Length

Description

Graphical view

Feature identifier

Molecule processing

Chain

1 – 575

575

Epsin-1

PRO_0000074515

Regions

Domain

12 – 144

133

ENTH

Repeat

183 – 202

20

UIM 1

Repeat

208 – 227

20

UIM 2

Repeat

233 – 252

20

UIM 3

Repeat

274 – 276

3

1

Repeat

294 – 296

3

2

Repeat

306 – 308

3

3

Repeat

319 – 321

3

4

Repeat

332 – 334

3

5

Repeat

349 – 351

3

6

Repeat

367 – 369

3

7

Repeat

377 – 379

3

8

Repeat

501 – 503

3

1

Repeat

517 – 519

3

2

Repeat

571 – 573

3

3

Region

274 – 379

106

8 X 3 AA repeats of D-P-W

Region

501 – 573

73

3 X 3 AA repeats of N-P-F

Motif

401 – 410

10

[DE]-X(1,2)-F-X-X-[FL]-X-X-X-R motif

Compositional bias

267 – 572

306

Ala/Gly/Pro-rich

Sites

Binding site

8

1

Phosphatidylinositol lipid headgroup

Binding site

11

1

Phosphatidylinositol lipid headgroup

Binding site

25

1

Phosphatidylinositol lipid headgroup

Binding site

30

1

Phosphatidylinositol lipid headgroup

Binding site

63

1

Phosphatidylinositol lipid headgroup

Binding site

73

1

Phosphatidylinositol lipid headgroup

Amino acid modifications

Modified residue

382

1

Phosphoserine By similarity

Modified residue

419

1

Phosphoserine By similarity

Modified residue

434

1

Phosphoserine By similarity

Modified residue

453

1

Phosphoserine By similarity

Modified residue

459

1

Phosphothreonine By similarity

Modified residue

463

1

Phosphothreonine By similarity

Modified residue

469

1

Phosphothreonine By similarity

Modified residue

493

1

Phosphothreonine By similarity

Experimental info

Mutagenesis

6

1

L → E, H or Q: Reduces lipid binding. Ref.10

Mutagenesis

8

1

R → A: Strongly reduces lipid binding. Ref.7

Mutagenesis

63

1

R → L: Strongly reduces lipid binding. Abolishes lipid binding; when associated with L-73. Ref.7 Ref.10

Mutagenesis

72

1

R → A: Abolishes ZNF145 binding. Ref.7

Mutagenesis

73

1

H → L: Abolishes lipid binding; when associated with L-63. Ref.10

Mutagenesis

76

1

K → A: Strongly reduces lipid binding. Ref.7

Mutagenesis

190

1

L → A: Abolishes mono-ubiquitination. Ref.8

Mutagenesis

257 – 259

3

LMD → AAA: Strongly reduces clathrin binding. Ref.6

Mutagenesis

260 – 263

4

LADV → AAAA: Strongly reduces clathrin binding. Ref.6

Secondary structure

1

.

.

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.

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575

Helix Strand Turn

Details...

Helix

2 – 11

10

Helix

19 – 27

9

Beta strand

30 – 33

4

Helix

37 – 46

10

Helix

50 – 64

15

Helix

68 – 70

3

Helix

71 – 87

17

Helix

90 – 98

9

Helix

100 – 104

5

Helix

105 – 108

4

Helix

120 – 135

16

Helix

137 – 155

19

Sequences

	Sequence		Length	Mass (Da)
	O88339-1 [UniParc]. Last modified November 1, 1998. Version 1. Checksum: D0B770F3B7AB5DDA	FASTA	575	60,158
	10 20 30 40 50 60 MSTSSLRRQM KNIVHNYSEA EIKVREATSN DPWGPSSSLM SEIADLTYNV VAFSEIMSMI 70 80 90 100 110 120 WKRLNDHGKN WRHVYKAMTL MEYLIKTGSE RVSQQCKENM YAVQTLKDFQ YVDRDGKDQG 130 140 150 160 170 180 VNVREKAKQL VALLRDEDRL REERAHALKT KEKLAQTATA SSAAVGSGPP PEAEQAWPQS 190 200 210 220 230 240 SGEEELQLQL ALAMSKEEAD QPPSCGPEDD VQLQLALSLS REEHDKEERI RRGDDLRLQM 250 260 270 280 290 300 AIEESKRETG GKEESSLMDL ADVFTTPAPP QASDPWGGPA SVPTAVPVAA AASDPWGAPA 310 320 330 340 350 360 VPPAADPWGG AAPTPASGDP WRPAAPTGPS VDPWGGTPAP AAGEGPTSDP WGSADGGAPV 370 380 390 400 410 420 SGPPSSDPWA PAPAFSDPWG GSPAKPSSNG TAVGGFDTEP DEFSDFDRLR TALPTSGSST 430 440 450 460 470 480 GELELLAGEV PARSPGAFDM SGVGGSLAES VGSPPPAATP TPTPPTRKTP ESFLGPNAAL 490 500 510 520 530 540 VDLDSLVSRP GPTPPGAKAS NPFLPSGAPA TGPSVTNPFQ PAPPATLTLN QLRLSPVPPV 550 560 570 PGAPPTYISP LGGGPGLPPM MPPGPPAPNT NPFLL

References

[1]	"Epsin is an EH-domain-binding protein implicated in clathrin-mediated endocytosis." Chen H., Fre S., Slepnev V.I., Capua M.R., Takei K., Butler M.H., Di Fiore P.P., De Camilli P. Nature 394:793-797(1998) [PubMed: 9723620] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [MRNA], FUNCTION, SUBCELLULAR LOCATION, TISSUE SPECIFICITY, INTERACTION WITH EPS15; AP2A1 AND AP2A2. Strain: Wistar. Tissue: Brain.
[2]	"A structural explanation for the binding of multiple ligands by the alpha-adaptin appendage domain." Owen D.J., Vallis Y., Noble M.E.M., Hunter J.B., Dafforn T.R., Evans P.R., McMahon H.T. Cell 97:805-815(1999) [PubMed: 10380931] [Abstract] Cited for: INTERACTION WITH AP2A2.
[3]	"The interaction of epsin and Eps15 with the clathrin adaptor AP-2 is inhibited by mitotic phosphorylation and enhanced by stimulation-dependent dephosphorylation in nerve terminals." Chen H., Slepnev V.I., Di Fiore P.P., De Camilli P. J. Biol. Chem. 274:3257-3260(1999) [PubMed: 9920862] [Abstract] Cited for: PHOSPHORYLATION, INTERACTION WITH AP-2.
[4]	"Crystal structure of the alpha appendage of AP-2 reveals a recruitment platform for clathrin-coat assembly." Traub L.M., Downs M.A., Westrich J.L., Fremont D.H. Proc. Natl. Acad. Sci. U.S.A. 96:8907-8912(1999) [PubMed: 10430869] [Abstract] Cited for: INTERACTION WITH AP2A2.
[5]	"The structure and function of the beta 2-adaptin appendage domain." Owen D.J., Vallis Y., Pearse B.M.F., McMahon H.T., Evans P.R. EMBO J. 19:4216-4227(2000) [PubMed: 10944104] [Abstract] Cited for: INTERACTION WITH AP2B1.
[6]	"Epsin binds to clathrin by associating directly with the clathrin-terminal domain. Evidence for cooperative binding through two discrete sites." Drake M.T., Downs M.A., Traub L.M. J. Biol. Chem. 275:6479-6489(2000) [PubMed: 10692452] [Abstract] Cited for: MUTAGENESIS OF 257-LEU--ASP-259 AND 260-LEU--VAL-263, SUBCELLULAR LOCATION, INTERACTION WITH CLATHRIN HEAVY CHAIN AND AP-2.
[7]	"Role of the ENTH domain in phosphatidylinositol-4,5-bisphosphate binding and endocytosis." Itoh T., Koshiba S., Kigawa T., Kikuchi A., Yokoyama S., Takenawa T. Science 291:1047-1051(2001) [PubMed: 11161217] [Abstract] Cited for: FUNCTION, INTERACTION WITH ZBTB16, SUBCELLULAR LOCATION, MUTAGENESIS OF ARG-8; ARG-63; ARG-72 AND LYS-76.
[8]	"A single motif responsible for ubiquitin recognition and monoubiquitination in endocytic proteins." Polo S., Sigismund S., Faretta M., Guidi M., Capua M.R., Bossi G., Chen H., De Camilli P., Di Fiore P.P. Nature 416:451-455(2002) [PubMed: 11919637] [Abstract] Cited for: MUTAGENESIS OF LEU-190, UBIQUITINATION.
[9]	"Epsin 1 undergoes nucleocytosolic shuttling and its eps15 interactor NH(2)-terminal homology (ENTH) domain, structurally similar to Armadillo and Heat repeats, interacts with the transcription factor promyelocytic leukemia Zn(2)+ finger protein (PLZF)." Hyman J., Chen H., Di Fiore P.P., De Camilli P., Brunger A.T. J. Cell Biol. 149:537-546(2000) [PubMed: 10791968] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (1.8 ANGSTROMS) OF 12-160, SUBCELLULAR LOCATION, INTERACTION WITH ZNF145.
[10]	"Curvature of clathrin-coated pits driven by epsin." Ford M.G.J., Mills I.G., Peter B.J., Vallis Y., Praefcke G.J.K., Evans P.R., McMahon H.T. Nature 419:361-366(2002) [PubMed: 12353027] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (1.7 ANGSTROMS) OF 1-158 IN COMPLEX WITH INOSITOL-1,4,5-TRIPHOSPHATE, FUNCTION, MUTAGENESIS OF LEU-6; ARG-63 AND HIS-73.
[11]	"Accessory protein recruitment motifs in clathrin-mediated endocytosis." Brett T.J., Traub L.M., Fremont D.H. Structure 10:797-809(2002) [PubMed: 12057195] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.0 ANGSTROMS) OF 375-381 IN COMPLEX WITH AP2A2.
+	Additional computationally mapped references.

Web resources

Protein Spotlight

The bubble's bend - Issue 42 of January 2004

Cross-references

Sequence databases

EMBL
GenBank
DDBJ

AF018261 mRNA. Translation: AAC33823.1.

IPI

IPI00208515.

RefSeq

NP_476477.1.

UniGene

Rn.30007

3D structure databases

PDBe
RCSB PDB
PDBj

Entry	Method	Resolution (Å)	Chain	Positions	PDBsum
1EDU	X-ray	1.80	A	12-160	[»]
1EYH	X-ray	1.56	A	15-158	[»]
1H0A	X-ray	1.70	A	1-158	[»]
1KY6	X-ray	2.00	P	375-381	[»]

DisProt

DP00251.

ModBase

Search...

Protein-protein interaction databases

STRING

O88339.

PTM databases

PhosphoSite

O88339.

Proteomic databases

PRIDE

O88339.

Genome annotation databases

Ensembl

ENSRNOT00000021286; ENSRNOP00000021286; ENSRNOG00000015753; Rattus norvegicus. [Genome view]

GeneID

117277.

KEGG

rno:117277.

UCSC

NM_057136. rat.

Organism-specific databases

CTD

117277.

RGD

619772. Epn1.

Phylogenomic databases

eggNOG

roNOG13892.

HOVERGEN

O88339.

InParanoid

O88339.

OMA

PTSDPWG.

OrthoDB

EOG9SXQZ5.

PhylomeDB

O88339.

Gene expression databases

ArrayExpress

O88339.

Genevestigator

O88339.

GermOnline

ENSRNOG00000015753. Rattus norvegicus.

Family and domain databases

InterPro

IPR008942. ENTH_VHS.
IPR013809. Epsin-like_N.
IPR001026. Epsin_N.
IPR003903. Ubiquitin-int_motif.
[Graphical view]

Gene3D

G3DSA:1.25.40.90. ENTH_VHS. 1 hit.

Pfam

PF01417. ENTH. 1 hit.
PF02809. UIM. 2 hits.
[Graphical view]

SMART

SM00273. ENTH. 1 hit.
SM00726. UIM. 3 hits.
[Graphical view]

PROSITE

PS50942. ENTH. 1 hit.
PS50330. UIM. 3 hits.
[Graphical view]

ProtoNet

Search...

Other Resources

NextBio

620217.

Entry information

Entry name

EPN1_RAT

Accession

Primary (citable) accession number: O88339

Entry history

Integrated into UniProtKB/Swiss-Prot:	January 16, 2004
Last sequence update:	November 1, 1998
Last modified:	January 19, 2010
This is version 91 of the entry and version 1 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)

Annotation project

HPI (Human Proteome Initiative)