Purification and functional characterization of β‐adrenergic receptor kinase expressed in insect cells

FEBS Letters

Sohlemann, Peter¹  Elce, John S.¹  Buchen, Claudia¹  Hekman, Mirko¹  Lohse, Martin J.¹ 

[1] Laboratory of Molecular Biology, University of Munich, Max-Planck-Institute of Biochemistry, 8033 Martinsried, Germany

关键词: β-Adrenergic receptor kinase;    β2-Adrenergic receptor;    Rhodopsin;    Receptor phosphorylation;    Baculovirus;    Spodoptera frugiperda cell;    βARK;    β-adrenergic receptor kinase;    CM;    carboxymethyl;    DEAE;    diethyl aminoethyl;    MOI;    multiplicity of infection (plaque forming units/cell);    PCR;    polymerase chain reaction;    SDS PAGE;    sodium dodecyl sulfate-polyacrylamide gel electrophoresis;   

DOI  :  10.1016/0014-5793(93)81532-5

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The β-adrenergic receptor kinase mediates agonist-dependent phosphorylation of β-adrenergic receptors, which is thought to represent the first step of homologous desensitization. We have expressed bovine and human βARK1 in Sf9 cells and purified them to apparent homogeneity in milligram quantities. The K _m-values of the enzyme were 3.8 μM for rhodopsin and 22 μM for ATP; the V _max-value was 9.9 mol phosphate/mol βARK/min. These data indicate that the two recombinant kinases were at least as active as preparations previously obtained from bovine brain. There were no differences in the functional activity of human and bovine βARK.

【 授权许可】

Unknown