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Regulation of Human Type V and Type VI Adenylyl Cyclases by G Protein βγ Subunits
Author(s) -
gao xianlong,
Sadana Rachna,
Dessauer Carmen,
Patel Tarun B
Publication year - 2006
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.20.5.lb111-c
Subject(s) - adenylyl cyclase , gs alpha subunit , protein subunit , adcy10 , forskolin , camp dependent pathway , adcy9 , gi alpha subunit , microbiology and biotechnology , transfection , g protein , chemistry , protein kinase a , adcy6 , sf9 , biochemistry , biology , in vitro , enzyme , receptor , recombinant dna , gene , spodoptera
We performed yeast two hybrid screening of mouse brain library using the N terminus of human type V adenylyl cyclase (ACV) as bait. We identified G protein β2 subunit as a candidate interacting protein of human ACV. Further, yeast two hybrid assay also showed β1 subunit interacted with the N terminus of human type VI adenylyl cyclase (ACVI). In vitro adenylyl cyclase assays using membranes from Sf9 cells expressing ACV and ACVI showed that βγ subunits enhance the activity of these enzymes when either Gsα or forskolin are present. However, when the N terminus of ACVI was deleted, activity of the enzyme could not be stimulated by G protein βγ subunits. Likewise, activity of the engineered, soluble form of ACVI, C1C2, also lacking the N‐terminus was not enhanced by βγ subunits. Additionally, transfection of the C terminus of β adrenal receptor kinase 1 to sequester βγ subunits, attenuated the ability of isoproterenol to increase cAMP in COS7 cells transfected to express human ACVI. These results suggest that the N termini of human ACV and ACVI are responsible for the interaction with and the regulation of their activities by βγ subunits both in‐vitro and in intact cells. Supported by NIH grant HL59679.