Characterization of RhoA-binding kinase ROKalpha implication of the pleckstrin homology domain in ROKalpha function using region-specific antibodies - PubMed (original) (raw)
. 2002 Apr 12;277(15):12680-8.
doi: 10.1074/jbc.M109839200. Epub 2002 Jan 28.
Affiliations
- PMID: 11815607
- DOI: 10.1074/jbc.M109839200
Free article
Characterization of RhoA-binding kinase ROKalpha implication of the pleckstrin homology domain in ROKalpha function using region-specific antibodies
Xiang-qun Chen et al. J Biol Chem. 2002.
Free article
Abstract
Rho-binding kinase alpha (ROKalpha) is a serine/threonine kinase with multiple functional domains involved in actomyosin assembly. It has previously been documented that the C terminus part of ROKalpha interacts with the N-terminal kinase domain and thereby regulates its catalytic activity. Here we used antibodies against different domains of ROKalpha and were able to reveal some structural aspects that are essential for the specific functions of ROKalpha. Antibodies against the kinase domain revealed that this part of the protein is highly complex and inaccessible. Further experiments confirmed that this domain could undergo inter- and intramolecular interactions in a complex manner, which regulates the kinase catalytic activity. Other antibodies that raised against the coiled-coil domain, Rho binding domain, and the pleckstrin homology (PH) domain were all effective in recognizing the native proteins in an immunoprecipitation assay. Only the anti-Rho binding domain antibodies could activate the kinase independent of RhoA. The PH antibodies had no apparent effects on the catalytic activity but were effective in blocking actomyosin assembly and cell contractility. Likewise, mutations of the PH domains can abrogate its dominant negative effects on actin morphology. The subsequent disruption of endogenous ROK localization to the actomyosin network by overexpressing the PH domain is supportive of a role of the PH domain of ROK in targeting the kinase to these structures.
Similar articles
- 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 XQ, Manser E, Lim L. Leung T, et al. Mol Cell Biol. 1996 Oct;16(10):5313-27. doi: 10.1128/MCB.16.10.5313. Mol Cell Biol. 1996. PMID: 8816443 Free PMC article. - p80 ROKalpha binding protein is a novel splice variant of CRMP-1 which associates with CRMP-2 and modulates RhoA-induced neuronal morphology.
Leung T, Ng Y, Cheong A, Ng CH, Tan I, Hall C, Lim L. Leung T, et al. FEBS Lett. 2002 Dec 18;532(3):445-9. doi: 10.1016/s0014-5793(02)03736-5. FEBS Lett. 2002. PMID: 12482610 - Dynamics of RhoA and ROKalpha translocation in single living cells.
Miyazaki K, Komatsu S, Ikebe M. Miyazaki K, et al. Cell Biochem Biophys. 2006;45(3):243-54. doi: 10.1385/CBB:45:3:243. Cell Biochem Biophys. 2006. PMID: 16845171 - RhoA-binding kinase alpha translocation is facilitated by the collapse of the vimentin intermediate filament network.
Sin WC, Chen XQ, Leung T, Lim L. Sin WC, et al. Mol Cell Biol. 1998 Nov;18(11):6325-39. doi: 10.1128/MCB.18.11.6325. Mol Cell Biol. 1998. PMID: 9774649 Free PMC article. - p160 RhoA-binding kinase ROKalpha induces neurite retraction.
Katoh H, Aoki J, Ichikawa A, Negishi M. Katoh H, et al. J Biol Chem. 1998 Jan 30;273(5):2489-92. doi: 10.1074/jbc.273.5.2489. J Biol Chem. 1998. PMID: 9446546
Cited by
- Druggable targets in the Rho pathway and their promise for therapeutic control of blood pressure.
Dee RA, Mangum KD, Bai X, Mack CP, Taylor JM. Dee RA, et al. Pharmacol Ther. 2019 Jan;193:121-134. doi: 10.1016/j.pharmthera.2018.09.001. Epub 2018 Sep 4. Pharmacol Ther. 2019. PMID: 30189292 Free PMC article. Review. - Evolving mechanisms of vascular smooth muscle contraction highlight key targets in vascular disease.
Liu Z, Khalil RA. Liu Z, et al. Biochem Pharmacol. 2018 Jul;153:91-122. doi: 10.1016/j.bcp.2018.02.012. Epub 2018 Feb 13. Biochem Pharmacol. 2018. PMID: 29452094 Free PMC article. Review. - ROCK as a therapeutic target for ischemic stroke.
Sladojevic N, Yu B, Liao JK. Sladojevic N, et al. Expert Rev Neurother. 2017 Dec;17(12):1167-1177. doi: 10.1080/14737175.2017.1395700. Epub 2017 Oct 30. Expert Rev Neurother. 2017. PMID: 29057688 Free PMC article. Review. - Apical Sarcomere-like Actomyosin Contracts Nonmuscle Drosophila Epithelial Cells.
Coravos JS, Martin AC. Coravos JS, et al. Dev Cell. 2016 Nov 7;39(3):346-358. doi: 10.1016/j.devcel.2016.09.023. Epub 2016 Oct 20. Dev Cell. 2016. PMID: 27773487 Free PMC article. - Structure of the Shroom-Rho Kinase Complex Reveals a Binding Interface with Monomeric Shroom That Regulates Cell Morphology and Stimulates Kinase Activity.
Zalewski JK, Mo JH, Heber S, Heroux A, Gardner RG, Hildebrand JD, VanDemark AP. Zalewski JK, et al. J Biol Chem. 2016 Dec 2;291(49):25364-25374. doi: 10.1074/jbc.M116.738559. Epub 2016 Oct 10. J Biol Chem. 2016. PMID: 27758857 Free PMC article.
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources