Expression, purification, and characterization of an enzymatically active truncated human rho-kinase I (ROCK I) domain expressed in Sf-9 insect cells |
| |
Authors: | Khandekar Sanjay S Yi Tracey Dul Ed Wright Lois L Chen Susan Scott Gilbert F Smith Gary K Lee Dennis Hu Erding Kirkpatrick Robert B |
| |
Institution: | Gene Expression and Protein Biochemistry, GlaxoSmithKline, King of Prussia, PA 19406, USA. sanjay.khandekar@gsk.com |
| |
Abstract: | Rho Kinase I (ROCK I) is a serine/threonine kinase that is involved in diverse cellular signaling. To further understand the physiological role of ROCK I and to identify and develop potent and selective inhibitors of ROCK I, we have overexpressed and purified a constitutively active dimeric human ROCK I (3-543) kinase domain using the Sf9-baculovirus expression system. In addition, using a limited proteolysis technique, we have identified a minimal functional subdomain of ROCK I that can be used in crystallization studies. The availability of multimilligram amounts of purified and well characterized functional human ROCK I kinase domains will be useful in screening and structural studies. |
| |
Keywords: | |
本文献已被 PubMed 等数据库收录! |
|