Binding of regulatory subunits of cyclic AMP-dependent protein kinase to cyclic CMP agarose.
The bacterial adenylyl cyclase toxins CyaA from Bordetella pertussis and edema factor from Bacillus anthracis as well as soluble guanylyl cyclase α(1)β(1) synthesize the cyclic pyrimidine nucleotide cCMP. These data raise the question to which effector proteins cCMP binds. Recently, we reported that cCMP activates the regulatory subunits RIα and RIIα of cAMP-dependent protein kinase. In this study, we used two cCMP agarose matrices as novel tools in combination with immunoblotting and mass spectrometry to identify cCMP-binding proteins. In agreement with our functional data, RIα and RIIα were identified as cCMP-binding proteins. These data corroborate the notion that cAMP-dependent protein kinase may serve as a cCMP target.
Institute of Pharmacology, Hannover Medical School, Hannover, Germany.
Andreas
A
Hammerschmidt
Bijon
B
Chatterji
Johannes
J
Zeiser
Anke
A
Schröder
Hans-Gottfried
HG
Genieser
Andreas
A
Pich
Volkhard
V
Kaever
Frank
F
Schwede
Sabine
S
Wolter
Roland
R
Seifert
D037361:Adenylate Cyclase Toxin / Q000378:metabolism; D054754:Cyclic AMP-Dependent Protein Kinase RIIalpha Subunit / Q000235:genetics / Q000378:metabolism*; D054756:Cyclic AMP-Dependent Protein Kinase RIalpha Subunit / Q000235:genetics / Q000378:metabolism*; D003563:Cyclic CMP / Q000737:chemistry / Q000378:metabolism*; D005786:Gene Expression Regulation; D006162:Guanylate Cyclase / Q000235:genetics / Q000378:metabolism; D057809:HEK293 Cells; D018922:HL-60 Cells; D006367:HeLa Cells; D006801:Humans; D015151:Immunoblotting; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D013058:Mass Spectrometry; D011485:Protein Binding; D012685:Sepharose / Q000737:chemistry*; D015398:Signal Transduction
D037361:Adenylate Cyclase Toxin; D054754:Cyclic AMP-Dependent Protein Kinase RIIalpha Subunit; D054756:Cyclic AMP-Dependent Protein Kinase RIalpha Subunit; D007527:Isoenzymes; D003563:Cyclic CMP; D012685:Sepharose; D006162:Guanylate Cyclase