Q62868 (ROCK2_RAT) 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
RhoA-binding kinase 2
p150 ROK-alpha
Short name=ROKalpha
p164 ROCK-2

Gene names

Name:

Rock2

Organism

Rattus norvegicus (Rat) [Reference proteome]

Taxonomic identifier

10116 [NCBI]

Taxonomic lineage

cellular organisms › Eukaryota › Opisthokonta › Metazoa › Eumetazoa › Bilateria › Deuterostomia › Chordata › Craniata › Vertebrata › Gnathostomata › Teleostomi › Euteleostomi › Sarcopterygii › Tetrapoda › Amniota › Mammalia › Theria › Eutheria › Euarchontoglires › Glires › Rodentia › Sciurognathi › Muroidea › Muridae › Murinae › Rattus

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. Plays a role in placental homeostasis during the perinatal period By similarity. Ref.1 Ref.2 Ref.3 Ref.8 Ref.9
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 CHORDC1, PPP1R12A, SORL1, EP300 and BRCA2 By similarity. Interacts with NPM1 and this interaction enhances its activity By similarity. Interacts with RHOA (activated by GTP), RHOB, RHOC and IRS1. Interacts with RAF1. Ref.1 Ref.2 Ref.4 Ref.5 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 Ref.8
Tissue specificity	Highly expressed in brain, lung, liver, skeletal muscle, kidney and testis. Ref.2
Domain	An interaction between Thr-414 and Asp-48 is essential for kinase activity and dimerization. Ref.7
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 Ref.2 Ref.7 Cleaved by granzyme B during apoptosis. This leads to constitutive activation of the kinase and membrane blebbing.
Involvement in disease	May play a role in hypertension. ROCK-inhibitors lower the blood pressure in spontaneous hypertensive, renal hypertensive and deoxycorticosterone acetate-induced hypertensive rats, but not in normal rats.
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.
Sequence caution	The sequence AAB37540.1 differs from that shown. Reason: Erroneous initiation. Translation N-terminally extended.

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 Reference proteome
Gene Ontology (GO)
Biological_process	Rho protein signal transduction Inferred from mutant phenotype PubMed 18332105. Source: MGI chromosome condensation Traceable author statement PubMed 12126956. Source: RGD
Cellular_component	cytoplasm Inferred from electronic annotation. Source: UniProtKB-SubCell 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 kinase activity Traceable author statement PubMed 12126956. Source: RGD metal ion binding Inferred from electronic annotation. Source: UniProtKB-KW phospholipid binding Inferred from electronic annotation. Source: InterPro 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_0000086627

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

Region

373 – 420

48

Interaction with NPM1 By similarity

Region

979 – 1047

69

RHOA binding By similarity

Coiled coil

439 – 1024

586

Potential

Coiled coil

1052 – 1131

80

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

Amino acid modifications

Modified residue

414

1

Phosphothreonine; by ROCK2 Ref.7

Modified residue

722

1

Phosphotyrosine; by SRC By similarity

Modified residue

1137

1

Phosphoserine By similarity

Experimental info

Mutagenesis

48

1

D → A: Loss of kinase activity; autophosphorylation and dimerization. Ref.7

Mutagenesis

121

1

K → A: Loss of kinase activity. Ref.2 Ref.7

Mutagenesis

121

1

K → M: Loss of autophosphorylation. Ref.2 Ref.7

Mutagenesis

414

1

T → A: Loss of kinase activity; autophosphorylation and dimerization. Ref.7

Mutagenesis

1170

1

W → A: Increased activity and autophosphorylation. Ref.7

Secondary structure

1

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

1388

Helix Strand Turn

Details...

Beta strand

1153 – 1158

6

Beta strand

1163 – 1166

4

Beta strand

1172 – 1178

7

Beta strand

1181 – 1186

6

Helix

1188 – 1192

5

Beta strand

1197 – 1200

4

Helix

1202 – 1204

3

Beta strand

1205 – 1210

6

Turn

1213 – 1215

3

Beta strand

1217 – 1219

3

Turn

1221 – 1223

3

Helix

1224 – 1226

3

Beta strand

1227 – 1232

6

Beta strand

1234 – 1236

3

Beta strand

1256 – 1258

3

Beta strand

1261 – 1266

6

Beta strand

1275 – 1281

7

Beta strand

1284 – 1286

3

Beta strand

1290 – 1296

7

Beta strand

1299 – 1301

3

Helix

1302 – 1307

6

Turn

1317 – 1319

3

Beta strand

1325 – 1330

6

Helix

1334 – 1347

14

Sequences

	Sequence		Length	Mass (Da)
	Q62868 [UniParc]. Last modified September 21, 2011. Version 2. Checksum: B23672CA8645067C	FASTA	1,388	160,387
	10 20 30 40 50 60 MSRPPPTGKM PGAPEAAAGD GAGAGRQRKL EALIRDPRSP INVESLLDGL NSLVLDLDFP 70 80 90 100 110 120 ALRKNKNIDN FLNRYEKIVK KIRGLQMKAE DYDVVKVIGR GAFGEVQLVR HKASQKVYAM 130 140 150 160 170 180 KLLSKFEMIK RSDSAFFWEE RDIMAFANSP WVVQLFCAFQ DDRYLYMVME 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 NSLCFPEDTE ISKHAKNLIC AFLTDREVRL GRNGVEEIKS ASFFKNDQWN 370 380 390 400 410 420 WDNIRETAAP VVPELSSDID SSNFDDIEDD KGDVETFPIP KAFVGNQLPF IGFTYFRENL 430 440 450 460 470 480 LLSDSPPCRE NDAIQTRKSE ESQEIQKKLY ALEEHLSSEV QAKEELEQKC KSINTRLEKT 490 500 510 520 530 540 AKELEEEITF RKNVESTLRQ LEREKALLQH KNAEYQRKAD HEADKKRNLE NDVNSLKDQL 550 560 570 580 590 600 EDLKKRNQSS QISTEKVNQL QKQLDEANAL LRTESDTAAR LRKTQAESSK QIQQLESNNR 610 620 630 640 650 660 DLQDKNCLLE TAKLKLEKEF INLQSALESE RRDRTHGSEI INDLQGRISG LEEDLKTGKT 670 680 690 700 710 720 LLAKVELEKR QLQEKLTDLE KEKSNMEIDM TYQLKVIQQS LEQEEAEHKT TKARLADKNK 730 740 750 760 770 780 IYESIEEAKS EAMKEMEKKL LEERSLKQKV ENLLLEAEKR CSILDCDLKQ SQQKLNELLK 790 800 810 820 830 840 QKDVLNEDVR NLTLKIEQET QKRCLMQNDL KMQTQQVNTL KMSEKQIKQE NNHLMEMKMN 850 860 870 880 890 900 LEKQNAELRK ERQDADGQMK ELQDQLEAEQ YFSTLYKTQV RELKEENEEK TKLCKELQQK 910 920 930 940 950 960 KQDLQDERDS LAAQLEITLT KADSEQLARS IAEEQYSDLE KEKIMKELEI KEMMARHKQE 970 980 990 1000 1010 1020 LTEKDATIAS LEETNRTLTS DVANLANEKE ELNNKLKDTQ EQLSKLKDEE ISAAAIKAQF 1030 1040 1050 1060 1070 1080 EKQLLTERTL KTQAVNKLAE IMNRKEPVKR GSDTDVRRKE KENRKLHMEL KSEREKLTQQ 1090 1100 1110 1120 1130 1140 MIKYQKELNE MQAQIAEESQ IRIELQMTLD SKDSDIEQLR SQLQALHIGM DSSSIGSGPG 1150 1160 1170 1180 1190 1200 DAEPDDGFPE SRLEGWLSLP VRNNTKKFGW VKKYVIVSSK KILFYDSEQD KEQSNPYMVL 1210 1220 1230 1240 1250 1260 DIDKLFHVRP VTQTDVYRAD AKEIPRIFQI LYANEGESKK EPEFPVEPVG EKSNYICHKG 1270 1280 1290 1300 1310 1320 HEFIPTLYHF PTNCEACMKP LWHMFKPPPA LECSRCHIKC HKDHMDKKEE IIAPCKVYYD 1330 1340 1350 1360 1370 1380 ISSAKNLLLL ANSTEEQQKW VSRLVKKIPK KPPAPDPFAR SSPRTSMKIQ QNQSIRRPSR QLAPNKPS

References

[1]	"A novel serine/threonine kinase binding the Ras-related RhoA GTPase which translocates the kinase to peripheral membranes." Leung T., Manser E., Tan L., Lim L. J. Biol. Chem. 270:29051-29054(1995) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [MRNA], FUNCTION, AUTOPHOSPHORYLATION, INTERACTION WITH RHOA, SUBCELLULAR LOCATION. Tissue: Brain.
[2]	"The p160 RhoA-binding kinase ROK alpha is a member of a kinase family and is involved in the reorganization of the cytoskeleton." Leung T., Chen X.-Q., Manser E., Lim L. Mol. Cell. Biol. 16:5313-5327(1996) [PubMed] [Europe PMC] [Abstract] Cited for: FUNCTION, AUTOPHOSPHORYLATION, INTERACTION WITH RHOA; RHOB AND RHOC, MUTAGENESIS OF LYS-121, TISSUE SPECIFICITY.
[3]	"Calcium sensitization of smooth muscle mediated by a Rho-associated protein kinase in hypertension." Uehata M., Ishizaki T., Satoh H., Ono T., Kawahara T., Morishita T., Tamakawa H., Yamagami K., Inui J., Maekawa M., Narumiya S. Nature 389:990-994(1997) [PubMed] [Europe PMC] [Abstract] Cited for: FUNCTION, ROLE IN HYPERTENSION, INHIBITION BY Y-27632.
[4]	"Active Rho kinase (ROK-alpha) associates with insulin receptor substrate-1 and inhibits insulin signaling in vascular smooth muscle cells." Begum N., Sandu O.A., Ito M., Lohmann S.M., Smolenski A. J. Biol. Chem. 277:6214-6222(2002) [PubMed] [Europe PMC] [Abstract] Cited for: INTERACTION WITH IRS1.
[5]	"Raf-1 regulates Rho signaling and cell migration." Ehrenreiter K., Piazzolla D., Velamoor V., Sobczak I., Small J.V., Takeda J., Leung T., Baccarini M. J. Cell Biol. 168:955-964(2005) [PubMed] [Europe PMC] [Abstract] Cited for: INTERACTION WITH RAF1.
[6]	"Direct cleavage of ROCK II by granzyme B induces target cell membrane blebbing in a caspase-independent manner." Sebbagh M., Hamelin J., Bertoglio J., Solary E., Breard J. J. Exp. Med. 201:465-471(2005) [PubMed] [Europe PMC] [Abstract] Cited for: CLEAVAGE BY GRANZYME B.
[7]	"The hydrophobic motif of ROCK2 requires association with the N-terminal extension for kinase activity." Couzens A.L., Saridakis V., Scheid M.P. Biochem. J. 419:141-148(2009) [PubMed] [Europe PMC] [Abstract] Cited for: DOMAIN, PHOSPHORYLATION AT THR-414, MUTAGENESIS OF ASP-48; LYS-121; THR-414 AND TRP-1170.
[8]	"ROCK isoform regulation of myosin phosphatase and contractility in vascular smooth muscle cells." Wang Y., Zheng X.R., Riddick N., Bryden M., Baur W., Zhang X., Surks H.K. Circ. Res. 104:531-540(2009) [PubMed] [Europe PMC] [Abstract] Cited for: FUNCTION, INTERACTION WITH PPP1R12A, SUBCELLULAR LOCATION.
[9]	"Chelerythrine perturbs lamellar actomyosin filaments by selective inhibition of myotonic dystrophy kinase-related Cdc42-binding kinase." Tan I., Lai J., Yong J., Li S.F., Leung T. FEBS Lett. 585:1260-1268(2011) [PubMed] [Europe PMC] [Abstract] Cited for: FUNCTION IN PHOSPHORYLATION OF MYL9/MLC2 AND PPP1R12A.
+	Additional computationally mapped references.

Cross-references

Sequence databases

EMBL
GenBank
DDBJ

U38481 mRNA. Translation: AAB37540.1. Different initiation.

IPI

IPI00565862.

RefSeq

NP_037154.2. NM_013022.2.

UniGene

Rn.88642.

3D structure databases

PDBe
RCSB PDB
PDBj

Entry	Method	Resolution (Å)	Chain	Positions	PDBsum
2ROV	NMR	-	A	1151-1351	[»]
2ROW	NMR	-	A	1237-1320	[»]

ModBase

Search...

Protein-protein interaction databases

MINT

MINT-4598028.

STRING

10116.ENSRNOP00000063773.

PTM databases

PhosphoSite

Q62868.

Proteomic databases

PaxDb

Q62868.

PRIDE

Q62868.

Protocols and materials databases

StructuralBiologyKnowledgebase

Search...

Genome annotation databases

GeneID

25537.

KEGG

rno:25537.

Organism-specific databases

CTD

9475.

RGD

3590. Rock2.

Phylogenomic databases

eggNOG

COG0515.

HOGENOM

HOG000017259.

HOVERGEN

HBG053111.

InParanoid

Q62868.

KO

K04514.

Gene expression databases

ArrayExpress

Q62868.

Genevestigator

Q62868.

GermOnline

ENSRNOG00000004496. Rattus norvegicus.

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

BindingDB

Q62868.

ChEMBL

CHEMBL5490.

EvolutionaryTrace

Q62868.

NextBio

607045.

Entry information

Entry name

ROCK2_RAT

Accession

Primary (citable) accession number: Q62868

Entry history

Integrated into UniProtKB/Swiss-Prot:	May 24, 2004
Last sequence update:	September 21, 2011
Last modified:	May 29, 2013
This is version 125 of the entry and version 2 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)

Annotation program

Chordata Protein Annotation Program