Q28021 (ROCK2_BOVIN) Reviewed, UniProtKB/Swiss-Prot

Names and origin

Protein names

Recommended name:
Rho-associated protein kinase 2
EC=2.7.11.1

Alternative name(s):
Rho-associated, coiled-coil-containing protein kinase 2
Rho-associated, coiled-coil-containing protein kinase II
Short name=ROCK-II
p164 ROCK-2

Gene names

Name:

ROCK2

Organism

Bos taurus (Bovine) [Reference proteome]

Taxonomic identifier

9913 [NCBI]

Taxonomic lineage

cellular organisms › Eukaryota › Opisthokonta › Metazoa › Eumetazoa › Bilateria › Deuterostomia › Chordata › Craniata › Vertebrata › Gnathostomata › Teleostomi › Euteleostomi › Sarcopterygii › Tetrapoda › Amniota › Mammalia › Theria › Eutheria › Laurasiatheria › Cetartiodactyla › Ruminantia › Pecora › Bovidae › Bovinae › Bos

Protein attributes

Sequence length	1388 AA.
Sequence status	Complete.
Protein existence	Evidence at protein level

General annotation (Comments)

Function	Protein kinase which is a key regulator of actin cytoskeleton and cell polarity. Involved in regulation of smooth muscle contraction, actin cytoskeleton organization, stress fiber and focal adhesion formation, neurite retraction, cell adhesion and motility via phosphorylation of ADD1, BRCA2, CNN1, EZR, DPYSL2, EP300, MSN, MYL9/MLC2, NPM1, RDX, PPP1R12A and VIM. Phosphorylates SORL1 and IRF4. Acts as a negative regulator of VEGF-induced angiogenic endothelial cell activation. Positively regulates the activation of p42/MAPK1-p44/MAPK3 and of p90RSK/RPS6KA1 during myogenic differentiation. Plays an important role in the timely initiation of centrosome duplication. Inhibits keratinocyte terminal differentiation. May regulate closure of the eyelids and ventral body wall through organization of actomyosin bundles. Plays a critical role in the regulation of spine and synaptic properties in the hippocampus. Ref.1 Ref.2 Ref.3 Ref.4 Ref.5 Ref.6 Ref.7
Catalytic activity	ATP + a protein = ADP + a phosphoprotein.
Cofactor	Magnesium By similarity.
Enzyme regulation	Activated by RHOA binding. Inhibited by Y-27632.
Subunit structure	Homodimer By similarity. Interacts with RHOA (activated by GTP), CHORDC1, IRS1, RHOB, RHOC, PPP1R12A, SORL1, EP300 and BRCA2 By similarity. Interacts with NPM1 and this interaction enhances its activity By similarity. Interacts with RAF1 By similarity. Ref.1 Ref.8
Subcellular location	Cytoplasm. Cell membrane; Peripheral membrane protein. Nucleus By similarity. Cytoplasm › cytoskeleton › centrosome By similarity. Note: Cytoplasmic, and associated with actin microfilaments and the plasma membrane. Ref.1
Tissue specificity	Highly expressed in whole brain and in cerebellum, and at lower levels in heart and lung. Detected at low levels in skeletal muscle, spleen, liver, kidney and pancreas. Ref.1
Domain	An interaction between Thr-414 and Asp-48 is essential for kinase activity and dimerization By similarity.
Post-translational modification	Autophosphorylated. Phosphorylation at Tyr-722 reduces its binding to RHOA and is crucial for focal adhesion dynamics. Dephosphorylation by PTPN11 stimulates its RHOA binding activity By similarity. Ref.1 Cleaved by granzyme B during apoptosis. This leads to constitutive activation of the kinase and membrane blebbing By similarity.
Sequence similarities	Belongs to the protein kinase superfamily. AGC Ser/Thr protein kinase family. Contains 1 AGC-kinase C-terminal domain. Contains 1 PH domain. Contains 1 phorbol-ester/DAG-type zinc finger. Contains 1 protein kinase domain. Contains 1 REM (Hr1) repeat.

Ontologies

Keywords
Cellular component	Cell membrane Cytoplasm Cytoskeleton Membrane Nucleus
Domain	Coiled coil Zinc-finger
Ligand	ATP-binding Magnesium Metal-binding Nucleotide-binding Zinc
Molecular function	Kinase Serine/threonine-protein kinase Transferase
PTM	Phosphoprotein
Technical term	3D-structure Complete proteome Direct protein sequencing Reference proteome
Gene Ontology (GO)
Biological_process	actin cytoskeleton organization Inferred from sequence or structural similarity. Source: AgBase intracellular signal transduction Inferred from electronic annotation. Source: InterPro
Cellular_component	cytosol Traceable author statement. Source: Reactome microtubule organizing center Inferred from electronic annotation. Source: UniProtKB-SubCell nucleus Inferred from electronic annotation. Source: UniProtKB-SubCell plasma membrane Inferred from electronic annotation. Source: UniProtKB-SubCell
Molecular_function	ATP binding Inferred from electronic annotation. Source: UniProtKB-KW metal ion binding Inferred from electronic annotation. Source: UniProtKB-KW phospholipid binding Inferred from electronic annotation. Source: InterPro protein kinase activity Inferred from direct assay PubMed 17015463. Source: MGI protein serine/threonine kinase activity Inferred from electronic annotation. Source: UniProtKB-KW
Complete GO annotation...

Sequence annotation (Features)

Feature key

Position(s)

Length

Description

Graphical view

Feature identifier

Molecule processing

Chain

1 – 1388

1388

Rho-associated protein kinase 2

PRO_0000086624

Regions

Domain

92 – 354

263

Protein kinase

Domain

357 – 425

69

AGC-kinase C-terminal

Repeat

475 – 559

85

REM

Domain

1150 – 1349

200

PH

Nucleotide binding

98 – 106

9

ATP By similarity

Zinc finger

1260 – 1315

56

Phorbol-ester/DAG-type

Region

363 – 784

422

Interaction with PPP1R12A By similarity

Region

373 – 420

48

Interaction with NPM1 By similarity

Region

979 – 1047

69

RHOA binding

Coiled coil

439 – 1025

587

Potential

Coiled coil

1053 – 1131

79

Potential

Sites

Active site

214

1

Proton acceptor By similarity

Binding site

121

1

ATP By similarity

Site

1131 – 1132

2

Cleavage; by granzyme B By similarity

Amino acid modifications

Modified residue

414

1

Phosphothreonine; by ROCK2 By similarity

Modified residue

722

1

Phosphotyrosine; by SRC By similarity

Modified residue

1137

1

Phosphoserine By similarity

Secondary structure

1

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

1388

Helix Strand Turn

Details...

Helix

30 – 33

4

Helix

43 – 57

15

Helix

62 – 64

3

Helix

66 – 82

17

Helix

83 – 85

3

Helix

89 – 91

3

Beta strand

92 – 100

9

Beta strand

105 – 111

7

Turn

112 – 114

3

Beta strand

117 – 124

8

Helix

125 – 130

6

Beta strand

135 – 137

3

Helix

138 – 146

9

Beta strand

155 – 160

6

Beta strand

162 – 169

8

Helix

177 – 183

7

Helix

188 – 206

19

Turn

207 – 209

3

Helix

217 – 219

3

Beta strand

220 – 222

3

Beta strand

228 – 230

3

Beta strand

241 – 247

7

Helix

254 – 256

3

Helix

259 – 263

5

Helix

264 – 266

3

Beta strand

269 – 272

4

Helix

274 – 289

16

Helix

299 – 307

9

Helix

309 – 312

4

Helix

323 – 332

10

Helix

336 – 338

3

Turn

340 – 343

4

Helix

346 – 349

4

Helix

352 – 354

3

Turn

361 – 363

3

Helix

364 – 366

3

Helix

408 – 410

3

Helix

979 – 1043

65

Sequences

	Sequence		Length	Mass (Da)
	Q28021 [UniParc]. Last modified November 1, 1996. Version 1. Checksum: CA83CE7D3860465D	FASTA	1,388	160,800
	10 20 30 40 50 60 MSRPPPTGKM PGAPEAVSGD GAGASRQRKL EALIRDPRSP INVESLLDGL NPLVLDLDFP 70 80 90 100 110 120 ALRKNKNIDN FLNRYEKIVK KIRGLQMKAE DYDVVKVIGR GAFGEVQLVR HKASQKVYAM 130 140 150 160 170 180 KLLSKFEMIK RSDSAFFWEE RDIMAFANSP WVVQLFCAFQ DDKYLYMVME YMPGGDLVNL 190 200 210 220 230 240 MSNYDVPEKW AKFYTAEVVL ALDAIHSMGL IHRDVKPDNM LLDKHGHLKL ADFGTCMKMD 250 260 270 280 290 300 ETGMVHCDTA VGTPDYISPE VLKSQGGDGY YGRECDWWSV GVFLFEMLVG DTPFYADSLV 310 320 330 340 350 360 GTYSKIMDHK NSLCFPEDAE ISKHAKNLIC AFLTDREVRL GRNGVEEIKQ HPFFKNDQWN 370 380 390 400 410 420 WDNIRETAAP VVPELSSDID SSNFDDIEDD KGDVETFPIP KAFVGNQLPF IGFTYYRENL 430 440 450 460 470 480 LLSDSPSCKE NDSIQSRKNE ESQEIQKKLY TLEEHLSTEI QAKEELEQKC KSVNTRLEKV 490 500 510 520 530 540 AKELEEEITL RKNVESTLRQ LEREKALLQH KNAEYQRKAD HEADKKRNLE NDVNSLKDQL 550 560 570 580 590 600 EDLKKRNQNS QISTEKVNQL QRQLDETNAL LRTESDTAAR LRKTQAESSK QIQQLESNNR 610 620 630 640 650 660 DLQDKNCLLE TAKLKLEKEF INLQSVLESE RRDRTHGSEI INDLQGRISG LEEDVKNGKI 670 680 690 700 710 720 LLAKVELEKR QLQERFTDLE KEKNNMEIDM TYQLKVIQQS LEQEETEHKA TKARLADKNK 730 740 750 760 770 780 IYESIEEAKS EAMKEMEKKL SEERTLKQKV ENLLLEAEKR CSILDCDLKQ SQQKINELLK 790 800 810 820 830 840 QKDVLNEDVR NLTLKIEQET QKRCLTQNDL KMQTQQVNTL KMSEKQLKQE NNHLLEMKMS 850 860 870 880 890 900 LEKQNAELRK ERQDADGQMK ELQDQLEAEQ YFSTLYKTQV RELKEECEEK TKLCKELQQK 910 920 930 940 950 960 KQELQDERDS LAAQLEITLT KADSEQLARS IAEEQYSDLE KEKIMKELEI KEMMARHKQE 970 980 990 1000 1010 1020 LTEKDATIAS LEETNRTLTS DVANLANEKE ELNNKLKEAQ EQLSRLKDEE ISAAAIKAQF 1030 1040 1050 1060 1070 1080 EKQLLTERTL KTQAVNKLAE IMNRKEPVKR GNDTDVRRKE KENRKLHMEL KSEREKLTQQ 1090 1100 1110 1120 1130 1140 MIKYQKELNE MQAQIAEESQ IRIELQMTLD SKDSDIEQLR SQLQALHIGL DSSSIGSGPG 1150 1160 1170 1180 1190 1200 DTEADDGFPE SRLEGWLSLP VRNNTKKFGW VKKYVIVSSK KILFYDSEQD KEQSNPYMVL 1210 1220 1230 1240 1250 1260 DIDKLFHVRP VTQTDVYRAD AKEIPRIFQI LYANEGESKK EQEFPVEPVG EKSNYICHKG 1270 1280 1290 1300 1310 1320 HEFIPTLYHF PTNCEACMKP LWHMFKPPPA LECRRCHIKC HKDHMDKKEE IIAPCKVYYD 1330 1340 1350 1360 1370 1380 ISSAKNLLLL ANSTEEQQKW VSRLVKKIPK KPPAPDPFAR SSPRTSMKIQ QNQSIRRPSR QLAPNKPS

References

[1]	"Rho-associated kinase, a novel serine/threonine kinase, as a putative target for small GTP binding protein Rho." Matsui T., Amano M., Yamamoto T., Chihara K., Nakafuku M., Ito M., Nakano T., Okawa K., Iwamatsu A., Kaibuchi K. EMBO J. 15:2208-2216(1996) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [MRNA], PROTEIN SEQUENCE OF 10-18; 30-44; 58-64; 133-140; 248-252; 291-305; 327-348; 350-360; 366-384; 392-400; 506-511; 527-535; 590-604; 633-652; 670-681; 829-842; 861-872; 913-921; 947-950; 979-988; 1066-1070; 1087-1091; 1114-1120; 1131-1141; 1145-1151; 1158-1166; 1182-1191; 1198-1218 AND 1318-1325, FUNCTION, INTERACTION WITH RHOA, PHOSPHORYLATION, TISSUE SPECIFICITY, SUBCELLULAR LOCATION.
[2]	"Phosphorylation and activation of myosin by Rho-associated kinase (Rho-kinase)." Amano M., Ito M., Kimura K., Fukata Y., Chihara K., Nakano T., Matsuura Y., Kaibuchi K. J. Biol. Chem. 271:20246-20249(1996) [PubMed] [Europe PMC] [Abstract] Cited for: FUNCTION.
[3]	"Phosphorylation of vimentin by Rho-associated kinase at a unique amino-terminal site that is specifically phosphorylated during cytokinesis." Goto H., Kosako H., Tanabe K., Yanagida M., Sakurai M., Amano M., Kaibuchi K., Inagaki M. J. Biol. Chem. 273:11728-11736(1998) [PubMed] [Europe PMC] [Abstract] Cited for: FUNCTION.
[4]	"Rho-kinase phosphorylates COOH-terminal threonines of ezrin/radixin/moesin (ERM) proteins and regulates their head-to-tail association." Matsui T., Maeda M., Doi Y., Yonemura S., Amano M., Kaibuchi K., Tsukita S., Tsukita S. J. Cell Biol. 140:647-657(1998) [PubMed] [Europe PMC] [Abstract] Cited for: FUNCTION.
[5]	"Phosphorylation of adducin by Rho-kinase plays a crucial role in cell motility." Fukata Y., Oshiro N., Kinoshita N., Kawano Y., Matsuoka Y., Bennett V., Matsuura Y., Kaibuchi K. J. Cell Biol. 145:347-361(1999) [PubMed] [Europe PMC] [Abstract] Cited for: FUNCTION.
[6]	"Identification of calponin as a novel substrate of Rho-kinase." Kaneko T., Amano M., Maeda A., Goto H., Takahashi K., Ito M., Kaibuchi K. Biochem. Biophys. Res. Commun. 273:110-116(2000) [PubMed] [Europe PMC] [Abstract] Cited for: FUNCTION.
[7]	"Phosphorylation of collapsin response mediator protein-2 by Rho-kinase. Evidence for two separate signaling pathways for growth cone collapse." Arimura N., Inagaki N., Chihara K., Menager C., Nakamura N., Amano M., Iwamatsu A., Goshima Y., Kaibuchi K. J. Biol. Chem. 275:23973-23980(2000) [PubMed] [Europe PMC] [Abstract] Cited for: FUNCTION.
[8]	"Parallel coiled-coil association of the RhoA-binding domain in Rho-kinase." Shimizu T., Ihara K., Maesaki R., Amano M., Kaibuchi K., Hakoshima T. J. Biol. Chem. 278:46046-46051(2003) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (1.8 ANGSTROMS) OF 979-1047 IN COMPLEX WITH RHOA, HOMODIMERIZATION.
+	Additional computationally mapped references.

Cross-references

Sequence databases

EMBL
GenBank
DDBJ

U36909 mRNA. Translation: AAC48567.1.

IPI

IPI00717918.

PIR

S70633.

RefSeq

NP_776877.1. NM_174452.2.

UniGene

Bt.120.

3D structure databases

PDBe
RCSB PDB
PDBj

Entry	Method	Resolution (Å)	Chain	Positions	PDBsum
1UIX	X-ray	1.80	A/B	979-1047	[»]
2F2U	X-ray	2.40	A/B	18-417	[»]
2H9V	X-ray	3.10	A	18-417	[»]

ProteinModelPortal

Q28021.

SMR

Q28021. Positions 27-417, 979-1045, 1151-1351.

ModBase

Search...

Protein-protein interaction databases

DIP

DIP-29038N.

IntAct

Q28021. 1 interaction.

Proteomic databases

PRIDE

Q28021.

Protocols and materials databases

StructuralBiologyKnowledgebase

Search...

Genome annotation databases

GeneID

282041.

KEGG

bta:282041.

Organism-specific databases

CTD

9475.

Phylogenomic databases

HOGENOM

HOG000017259.

HOVERGEN

HBG053111.

InParanoid

Q28021.

KO

K04514.

OrthoDB

EOG4PZJ5T.

Enzyme and pathway databases

Reactome

REACT_115433. Developmental Biology.

Family and domain databases

Gene3D

2.30.29.30. 2 hits.

InterPro

IPR000961. AGC-kinase_C.
IPR011072. HR1_rho-bd.
IPR011009. Kinase-like_dom.
IPR011993. PH_like_dom.
IPR017892. Pkinase_C.
IPR001849. Pleckstrin_homology.
IPR002219. Prot_Kinase_C-like_PE/DAG-bd.
IPR000719. Prot_kinase_cat_dom.
IPR017441. Protein_kinase_ATP_BS.
IPR020684. Rho-assoc_coiled-coil_kin.
IPR015008. Rho-bd_dom.
IPR002290. Ser/Thr_dual-sp_kinase_dom.
IPR008271. Ser/Thr_kinase_AS.
[Graphical view]

PANTHER

PTHR22988:SF3. PTHR22988:SF3. 1 hit.

Pfam

PF02185. HR1. 1 hit.
PF00169. PH. 1 hit.
PF00069. Pkinase. 1 hit.
PF00433. Pkinase_C. 1 hit.
PF08912. Rho_Binding. 1 hit.
[Graphical view]

PIRSF

PIRSF037568. Rho_kinase. 1 hit.

SMART

SM00109. C1. 1 hit.
SM00233. PH. 1 hit.
SM00133. S_TK_X. 1 hit.
SM00220. S_TKc. 1 hit.
[Graphical view]

SUPFAM

SSF56112. Kinase_like. 1 hit.

PROSITE

PS51285. AGC_KINASE_CTER. 1 hit.
PS50003. PH_DOMAIN. 1 hit.
PS00107. PROTEIN_KINASE_ATP. 1 hit.
PS50011. PROTEIN_KINASE_DOM. 1 hit.
PS00108. PROTEIN_KINASE_ST. 1 hit.
PS00479. ZF_DAG_PE_1. False negative.
PS50081. ZF_DAG_PE_2. 1 hit.
[Graphical view]

ProtoNet

Search...

Other

EvolutionaryTrace

Q28021.

NextBio

20805903.

Entry information

Entry name

ROCK2_BOVIN

Accession

Primary (citable) accession number: Q28021

Entry history

Integrated into UniProtKB/Swiss-Prot:	May 24, 2004
Last sequence update:	November 1, 1996
Last modified:	May 1, 2013
This is version 123 of the entry and version 1 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)

Annotation program

Chordata Protein Annotation Program