Rho Associated Coiled Coil containing Protein kinase 2 from MyBioSource.com

Rho-kinase (ROCK) is one of the best-characterized effectors of small GTPase RhoA and belongs to the AGC (protein kinase A/protein kinase G/protein kinase C) family of serine/threonine kinases. ROCK family contains two members, ROCK1 and ROCK2; they share 65% homology and 92% overall identity in the kinase domain. ROCK family of kinases promote actin-Myosin mediated contractile force generated by phosphorylation of downstream targets that include myosin binding subunit of myosin light chain (MLC) phosphatase 1 (MYPT1), myosin light chain 2 (MLC2), and LIM kinase (1). These interactions led rearrangements in actin cytoskeletal organization, stress fiber formation and smooth muscle cell contraction. Both kinases contain a catalytic kinase domain at the N-terminus, followed by a central coiled-coil domain, including a Rho-binding domain and a C-terminal pleckstrin-homology (PH) domain, with an internal cysteine-rich domain (2). Both ROCK1 and ROCK2, kinases contain a catalytic kinase domain at the N-terminus, followed by a central coiled-coil domain, including a Rho-binding domain and a C-terminal pleckstrin-homology (PH) domain, with an internal cysteine-rich domain. The two most commonly used inhibitors for these two kinases are Y2763217 and fasudil,18 target the ATP-dependent kinase domain, therefore inhibit ROCK1 and ROCK2 with equal potency with non-selective effects (3).

The two isoforoms of the Rho-kinases, ROCK1 and ROCK2are very similar and somewhat redundant. ROCK2 has been implicated in numerous pathologies and the current focus is on understanding the molecular events that couple ROCK2 activity to biological function. Deletion of ROCK2 in cardiomyocytes leads to increased expression of four-and-a-half LIM-only protein-2 (FHL2) and FHL2-mediated inhibition of serum response factor (SRF) and extracellular signal-regulated mitogen-activated protein kinase (ERK, 4). The phosphorylation of interferon regulated factor 4 (IRF4) by ROCK2 regulated the production of inflammatory interleukins like IL17 and IL21suggesting ROCK2 could be an important therapeutic target for the treatment of autoimmune disorders (5).

ROCK1-selective antibodies were generated against peptides taken from the human ROCK2 protein. The ROCK2-selective antibodies are affinity purified on an immobilized antigen based affinity matrix, the isolated antibodies were then stabilized in antibody stabilization buffer for long-term storage. The ROCK2-selective antibodies are fully characterized for applications in western blotting and ELISA at the recommended dilutions