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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
7
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pubmed:dateCreated |
2000-4-24
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pubmed:abstractText |
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.
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pubmed:grant |
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pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/10737781-10570143,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10737781-10737782,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10737781-1522899,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10737781-1730692,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10737781-1956325,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10737781-2071581,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10737781-2962193,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10737781-6212740,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10737781-690139,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10737781-7797516,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10737781-7836439,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10737781-8226899,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10737781-8876162,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10737781-9050844,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10737781-942051,
http://linkedlifedata.com/resource/pubmed/commentcorrection/10737781-9759500
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
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pubmed:status |
MEDLINE
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pubmed:month |
Mar
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pubmed:issn |
0027-8424
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:day |
28
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pubmed:volume |
97
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
3004-9
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pubmed:dateRevised |
2009-11-19
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pubmed:meshHeading |
pubmed-meshheading:10737781-Adenosine Triphosphate,
pubmed-meshheading:10737781-Animals,
pubmed-meshheading:10737781-COS Cells,
pubmed-meshheading:10737781-Cattle,
pubmed-meshheading:10737781-Cell Line,
pubmed-meshheading:10737781-Chromatography, Liquid,
pubmed-meshheading:10737781-Cloning, Molecular,
pubmed-meshheading:10737781-Electrophoresis, Polyacrylamide Gel,
pubmed-meshheading:10737781-Eye Proteins,
pubmed-meshheading:10737781-G-Protein-Coupled Receptor Kinase 1,
pubmed-meshheading:10737781-Humans,
pubmed-meshheading:10737781-Hydrogen-Ion Concentration,
pubmed-meshheading:10737781-Protein Kinases,
pubmed-meshheading:10737781-Protein Prenylation,
pubmed-meshheading:10737781-Recombinant Proteins,
pubmed-meshheading:10737781-Retina
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pubmed:year |
2000
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pubmed:articleTitle |
Rhodopsin kinase: expression in mammalian cells and a two-step purification.
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pubmed:affiliation |
Departments of Biology and Chemistry, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, MA 02139, USA.
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pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.,
Research Support, Non-U.S. Gov't
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