Autophosphorylation at the regulatory beta subunit reflects the supramolecular organization of protein kinase CK2.
Among the features of protein kinase CK2, autophosphorylation at its beta-subunit(s) upon incubation with ATP/Mg++ was early detected as a rapid and stoichiometric event occurring through an intramolecular mechanism as judged from kinetic analyses. The autophosphorylation site was mapped to Ser2 and, to a lesser extent, Ser3 both fulfilling the CK2 consensus sequence (MSSSEEV). The crystal structure of the heterotetrameric holoenzyme, however, is not compatible with an intramolecular autophosphorylation of the N-terminal stretch of either of the two beta subunits. Here we show that efficient "intramolecular" autophosphorylation of the beta subunit is crucially dependent on the formation of oligomers composed by several holoenzyme heterotetrameric protomers. Increasing ionic strength of the incubation medium promoting dissociation of the supramolecular oligomers abrogates beta subunit autophosphorylation, although CK2 catalytic activity, as judged from the phosphorylation of exogenous substrates, is still quite evident. These findings, in conjunction with graphic modelization, support the view that CK2 autophosphorylation at its beta subunits takes place through an "intraoligomeric" mechanism where the beta subunits of a protomer are phosphorylated by the catalytic subunits of another adjacent protomer. It appears therefore that in vivo beta autophosphorylation is symptomatic of supramolecular CK2 oligomers.
Pagano
Mario A
MA
Dipartimento di Chimica Biologica, Universittà di Padova, viale G. Colombo 3, 35121 Padova, Italy.
Sarno
Stefania
S
Poletto
Giorgia
G
Cozza
Giorgio
G
Pinna
Lorenzo A
LA
Meggio
Flavio
F
D000818:Animals; D047390:Casein Kinase II / Q000737:chemistry*; D004789:Enzyme Activation; D020035:Holoenzymes / Q000737:chemistry; D008958:Models, Molecular; D010766:Phosphorylation; D021122:Protein Subunits / Q000737:chemistry; D051381:Rats; D012694:Serine / Q000737:chemistry
D020035:Holoenzymes; D021122:Protein Subunits; D012694:Serine; D047390:Casein Kinase II