Literature DB >> 10737781

Rhodopsin kinase: expression in mammalian cells and a two-step purification.

C Bruel1, K Cha, P J Reeves, E Getmanova, H G Khorana.   

Abstract

A suitable system for expression of the rhodopsin kinase (RK) gene and its mutants is needed for structure-function studies of RK. Previously, investigation of the baculovirus system showed satisfactory production of RK, but posttranslational isoprenylation was deficient. We now report on a comparative study of expression of the RK gene in yeast (Pichia pastoris), COS-1 cells and in an HEK293 stable cell line. Expression in COS-1 cells, by using pCMV5 vector, is the most satisfactory. A two-step procedure for purification of the expressed enzyme with an N-terminal histidine tag has been developed. The purified enzyme has correct posttranslational modifications and shows a somewhat broader pH vs. catalytic activity profile than the wild-type enzyme.

Entities:  

Mesh：

Substances：

Year:  2000        PMID： 10737781      PMCID： PMC16182          DOI： 10.1073/pnas.97.7.3004

Source DB:  PubMed          Journal:  Proc Natl Acad Sci U S A        ISSN： 0027-8424            Impact factor:   11.205


  17 in total

1.  Mechanism of rhodopsin kinase activation.

Authors:  K Palczewski; J Buczyłko; M W Kaplan; A S Polans; J W Crabb
Journal:  J Biol Chem       Date:  1991-07-15       Impact factor: 5.157

2.  Protein kinase catalytic domain sequence database: identification of conserved features of primary structure and classification of family members.

Authors:  S K Hanks; A M Quinn
Journal:  Methods Enzymol       Date:  1991       Impact factor: 1.600

Review 3.  G protein-coupled receptor kinases.

Authors:  J A Pitcher; N J Freedman; R J Lefkowitz
Journal:  Annu Rev Biochem       Date:  1998       Impact factor: 23.643

4.  A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.

Authors:  M M Bradford
Journal:  Anal Biochem       Date:  1976-05-07       Impact factor: 3.365

5.  Expression of a synthetic bovine rhodopsin gene in monkey kidney cells.

Authors:  D D Oprian; R S Molday; R J Kaufman; H G Khorana
Journal:  Proc Natl Acad Sci U S A       Date:  1987-12       Impact factor: 11.205

6.  NMR spectroscopy in studies of light-induced structural changes in mammalian rhodopsin: applicability of solution (19)F NMR.

Authors:  J Klein-Seetharaman; E V Getmanova; M C Loewen; P J Reeves; H G Khorana
Journal:  Proc Natl Acad Sci U S A       Date:  1999-11-23       Impact factor: 11.205

7.  Preparation of antibodies to rhodopsin and the large protein of rod outer segments.

Authors:  D S Papermaster
Journal:  Methods Enzymol       Date:  1982       Impact factor: 1.600

8.  Light-dependent phosphorylation of rhodopsin. Purification and properties of rhodopsin kinase.

Authors:  H Shichi; R L Somers
Journal:  J Biol Chem       Date:  1978-10-10       Impact factor: 5.157

9.  Isoprenylation in regulation of signal transduction by G-protein-coupled receptor kinases.

Authors:  J Inglese; W J Koch; M G Caron; R J Lefkowitz
Journal:  Nature       Date:  1992-09-10       Impact factor: 49.962

10.  Isoprenylation of a protein kinase. Requirement of farnesylation/alpha-carboxyl methylation for full enzymatic activity of rhodopsin kinase.

Authors:  J Inglese; J F Glickman; W Lorenz; M G Caron; R J Lefkowitz
Journal:  J Biol Chem       Date:  1992-01-25       Impact factor: 5.157
View more
  4 in total

1.  Rhodopsin kinase: two mAbs binding near the carboxyl terminus cause time-dependent inactivation.

Authors:  C Bruel; K Cha; L Niu; P J Reeves; H G Khorana
Journal:  Proc Natl Acad Sci U S A       Date:  2000-03-28       Impact factor: 11.205

2.  Structure and function in rhodopsin: a tetracycline-inducible system in stable mammalian cell lines for high-level expression of opsin mutants.

Authors:  Philip J Reeves; Jong-Myoung Kim; H Gobind Khorana
Journal:  Proc Natl Acad Sci U S A       Date:  2002-10-07       Impact factor: 11.205

3.  Modulation of mouse rod response decay by rhodopsin kinase and recoverin.

Authors:  Ching-Kang Chen; Michael L Woodruff; Frank S Chen; Yenlin Chen; Marianne C Cilluffo; Daniel Tranchina; Gordon L Fain
Journal:  J Neurosci       Date:  2012-11-07       Impact factor: 6.167

4.  Structural insights into the inhibition mechanism of human sterol O-acyltransferase 1 by a competitive inhibitor.

Authors:  Chengcheng Guan; Yange Niu; Si-Cong Chen; Yunlu Kang; Jing-Xiang Wu; Koji Nishi; Catherine C Y Chang; Ta-Yuan Chang; Tuoping Luo; Lei Chen
Journal:  Nat Commun       Date:  2020-05-18       Impact factor: 14.919
  4 in total

北京卡尤迪生物科技股份有限公司 © 2022-2023.