Rho-Kinases

Rho-kinases (aka Rho-associated coiled-coil kinases or ROCKs) are serine/threonine kinases that are so called because they are thought to be activated by RhoA GTPases. They are modulators of processes involving cytoskeletal rearrangement, such as focal adhesion formation, cell motility and tumor cell invasion. ROCK inhibitors such as Y-27632 (Cat. No. 1254) are widely used in stem cell culture, for maintenance and differentiation protocols.

Products
Background
Literature (6)
Gene Data

Inhibitors

Cat. No. Product Name / Activity
4927 AS 1892802
Potent ROCK inhibitor; orally bioavailable
7163 Chroman 1
Highly potent and selective ROCK 2 inhibitor; improves cell survival after cryogeneisis
0541 Fasudil hydrochloride
Inhibitor of cyclic nucleotide dependent- and Rho-kinases
4009 GSK 269962
Potent and selective ROCK inhibitor
3726 GSK 429286
Selective Rho-kinase (ROCK) inhibitor
2414 H 1152 dihydrochloride
Selective Rho-kinase (ROCK) inhibitor
2485 Glycyl-H 1152 dihydrochloride
Selective Rho-kinase (ROCK) inhibitor. More selective analog of H 1152 dihydrochloride (Cat. No. 2414)
2415 HA 1100 hydrochloride
Cell-permeable, selective Rho-kinase inhibitor
5182 OXA 06 dihydrochloride
Potent ROCK inhibitor
5061 RKI 1447 dihydrochloride
Potent and selective ROCK inhibitor; antitumor
4118 SB 772077B dihydrochloride
Potent Rho-kinase inhibitor; vasodilator
3667 SR 3677 dihydrochloride
Potent, selective Rho-kinase (ROCK) inhibitor
4961 TC-S 7001
Potent and highly selective ROCK inhibitor; orally active
3845 Thiazovivin
ROCK inhibitor ; improves the efficiency of fibroblast reprogramming and induction of iPSCs
1254 Y-27632 dihydrochloride
Selective ROCK inhibitor; inhibits ROCK1 (p160 ROCK) and ROCK2; stem cell freezing (cryopreservation) media component

There are two isoforms of ROCK, ROCK1 (aka ROKβ or p160 ROCK) and ROCK2 (aka ROKα), which share a high degree of structural identity in their kinase domains (92%). The two Rho-kinase isoforms have different patterns of tissue expression; ROCK2 expression is highest in the brain and in muscle, whereas ROCK1 has a ubiquitous tissue distribution. Subcellularly, ROCK2 is found in the cytosol and localizes to the cleavage furrow during cytokinesis. ROCK1 is thought to colocalize with the centrosomes having an important role in cell cycle control.

ROCKs consist of a central coiled-coil of around 700 amino acids, forming a bridge between the N-terminal AGC kinase domain and the C-terminal split-PH domain, which has a zinc-finger cysteine-rich domain (CRD, also referred to as a C1 domain) spliced into it. ROCK was previously thought to be a downstream effector molecule of the Rho-GTPase, RhoA; ROCK activity was thought to be increased by binding of RhoA. However, more recent research suggests that ROCK is not bound or activated by RhoA, but exhibits constitutive activity, which is regulated by the length of the coiled-coil segment.

ROCK has a role in cell-cell adhesion via an E-cadherin dependent mechanism; it is thought that E-cadherin mediated cell-cell interaction regulates ROCK activity. Stem cells in culture are vulnerable to apoptosis as a result of single cell dissociation during routine passage. This is likely due to loss of cell-cell interaction and hyperactivation of ROCK signaling. It has been found that ROCK inhibitors such as Y-27632 (Cat. No. 1254) can reduce dissociation-induced apoptosis of embryonic stem cells (ESCs), improving cell survival and colony formation; inhibitors of other kinases are ineffective. Y-27632, is also key component of several organoid and stem cell protocols. ROCK inhibitors are also found to improve survival of stem cells undergoing cryopreservation when added to the cryopreservation medium and/or to the post-thaw medium.


Cryopreservation of Stem Cells

Schematic showing use of ROCK inhibitor Y-27632 in cryopreservation of stem cells

Figure 1: Schematic highlighting the use of ROCK inhibitor Y-27632 to improve survival of human pluripotent stem cells undergoing cryopreservation.

Adapted from Liu and Chen (2014) Cryopreservation of human pluripotent stem cells in defined medium. Curr.Protoc.Stem.Cell Biol., 31, 1C.17.1-13.


ROCKs phosphorylate a number of proteins involved in actin filament assembly and contraction, such as LIM Kinases, Myosin Light Chain and ERM proteins. Through these actions ROCKs have an important role in cell migration by controlling actin polymerization and cellular contractility. Other downstream targets of the Rho kinases include signal transduction molecules such as IRS-1 and PI 3-K/AKT.

External sources of pharmacological information for Rho-Kinases :

Literature for Rho-Kinases

Tocris offers the following scientific literature for Rho-Kinases to showcase our products. We invite you to request* or download your copy today!

*Please note that Tocris will only send literature to established scientific business / institute addresses.


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Rho-Kinases Gene Data

Gene Species Gene Symbol Gene Accession No. Protein Accession No.
ROCKI (ROKβ, p160ROCK) Human ROCK1 NM_005406 Q13464
Mouse Rock1 NM_009071 P70335
Rat Rock1 NM_031098 Q63644
ROCKII (ROKα) Human ROCK2 NM_004850 O75116
Mouse Rock2 NM_009072 P70336
Rat Rock2 NM_013022 Q62868