Adenylyl cyclase (AC) was extracted from human myometrium with either non-ionic (Lubrol-PX or Triton X-100) or zwitterionic (3-[3-cholamidopropyl)dimethylammonio]-1-propanesulfonate, CHAPS) detergents. The soluble enzyme was stimulated by forskolin, a hydrophobic activator, in the presence of Mg2+ indicating that the catalytic subunit had not been damaged after solubilization. The enzyme was also activated by 5′-guanylyl imidodiphosphate (Gpp(NH)p) showing that the catalytic unit was not separated from stimulatory guanine nucleotide binding protein (Gs) during the extraction. Both activators showed different effects on the stimulatory efficacy and potency of AC activity solobulized with detergents. Gel filtration of Lubrol-PX and CHAPS extracts over a Sepharose CL-2B column partially resolved AC and its complexes. The chromatographic profile for Lubrolsolubilized AC presented a main peak of about 200 kDa whereas CHAPS-solubilized AC showed a dominant peak of about 1100 kDa. The heterodisperse peaks obtained revealed that the catalytic AC subunit was not separated from Gs proteins after gel filtration, and that AC could be associated with other cellular proteins. When Lubrol extract was submitted to anionic-exchange chromatography, the enzyme was purified about 7.5 fold (enzymatic activity of 48.1 pmol/min/mg of protein). The catalytic subunit was co-eluted with both AC-activating proteins Gαs large (52.2 kDa) and Gαs small (48.7 kDa). This is the first demonstration of the stable physical association of AC with both αs subunits of G proteins in human myometrium.
Article navigation
Research Article|
August 01 2003
Solubilization of Adenylyl Cyclase from Human Myometrium in a αS-Coupled Form
Biosci Rep (2003) 23 (4): 175-186.
Citation
Ana M. Bajo, Juan C. Prieto, Pedro Valenzuela, Pilar Martinez, Luis G. Guijarro; Solubilization of Adenylyl Cyclase from Human Myometrium in a αS-Coupled Form. Biosci Rep 1 August 2003; 23 (4): 175–186. doi: https://doi.org/10.1023/B:BIRE.0000007691.17175.d9
Download citation file:
Close
Sign in
Don't already have an account? Register
Sign in to your personal account
You could not be signed in. Please check your email address / username and password and try again.
Biochemical Society Member Sign in
Sign InSign in via your Institution
Sign in via your Institution
0
Views
0
Citations
Cited By
Related Articles
Synthesis and degradation of cAMP in Giardia lamblia : possible role and characterization of a nucleotidyl cyclase with a single cyclase homology domain
Biochem J (November, 2017)
Regulation and organization of adenylyl cyclases and cAMP
Biochem J (November, 2003)
Adenylyl cyclase 6 is required for maintaining acid–base homeostasis
Clin Sci (Lond) (August, 2018)
Coxsackievirus B3 entry into the host cell interferes with G-protein-mediated transmembrane signalling
Biosci Rep (August, 1994)