title: Biochemical and three-dimensional-structural study of the specific inhibition of protein kinase CK2 by [5-oxo-5,6-dihydroindolo-(1,2-a)quinazolin-7-yl]acetic acid (IQA).
abstract: IQA [[5-oxo-5,6-dihydro-indolo(1,2-a)quinazolin-7-yl]acetic acid] is a novel ATP/GTP site-directed inhibitor of CK2 ('casein kinase 2'), a pleiotropic and constitutively active protein kinase whose activity is abnormally high in transformed cells. The K (i) value of IQA (0.17 microM) is lower than those of other CK2 inhibitors reported so far. Tested at 10 microM concentration in the presence of 100 microM ATP, IQA almost suppresses CK2 activity in vitro, whereas it is ineffective or weakly effective on a panel of 44 protein kinases and on phosphoinositide 3-kinase. In comparison, other CK2 inhibitors, notably apigenin and quercetin, are more promiscuous. The in vivo efficacy of IQA has been assessed by using the fact that treatment of Jurkat cells with IQA inhibits endogenous CK2 in a dose-dependent manner. IQA has been co-crystallized with maize CK2alpha, which is >70% identical with its human homologue, and the structure of the complex has been determined at 1.68 A (1 A=0.1 nm) resolution. The inhibitor lies in the same plane occupied by the purine moiety of ATP with its more hydrophobic side facing the hinge region. Major contributions to the interaction are provided by hydrophobic forces and non-polar interactions involving the aromatic portion of the inhibitor and the hydrophobic residues surrounding the ATP-binding pocket, with special reference to the side chains of V53 (Val53), I66, M163 and I174. Consequently, mutants of human CK2alpha in which either V66 (the homologue of maize CK2alpha I66) or I174 is replaced by alanine are considerably less sensitive to IQA inhibition when compared with wild-type. These results provide new tools for deciphering the enigmatic role of CK2 in living cells and may pave the way for the development of drugs depending on CK2 activity.
authors.affiliation: Venetian Institute for Molecular Medicine (VIMM), University of Padova, 35121 Padova, Italy.
authors.forename: Stefania
authors.initials: S
authors.lastname: Sarno
authors.forename: Erika
authors.initials: E
authors.lastname: de Moliner
authors.forename: Maria
authors.initials: M
authors.lastname: Ruzzene
authors.forename: Mario A
authors.initials: MA
authors.lastname: Pagano
authors.forename: Roberto
authors.initials: R
authors.lastname: Battistutta
authors.forename: Jenny
authors.initials: J
authors.lastname: Bain
authors.forename: Doriano
authors.initials: D
authors.lastname: Fabbro
authors.forename: Joseph
authors.initials: J
authors.lastname: Schoepfer
authors.forename: Matthew
authors.initials: M
authors.lastname: Elliott
authors.forename: Pascal
authors.initials: P
authors.lastname: Furet
authors.forename: Flavio
authors.initials: F
authors.lastname: Meggio
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Zanotti
authors.forename: Lorenzo A
authors.initials: LA
authors.lastname: Pinna
mesh_terms: D000085:Acetates / Q000737:chemistry*; D000818:Animals; D047390:Casein Kinase II; D002460:Cell Line; D018360:Crystallography, X-Ray; D004791:Enzyme Inhibitors / Q000737:chemistry*; D006801:Humans; D019169:Jurkat Cells; D015394:Molecular Structure; D010940:Plant Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors* / Q000737:chemistry*; D020836:Protein Structure, Quaternary / Q000187:drug effects; D011799:Quinazolines / Q000737:chemistry*; D051381:Rats; D003313:Zea mays / Q000201:enzymology
chemical_list: C475515:(5-oxo-5,6-dihydroindolo(1,2-a)quinazolin-7-yl)acetic acid; D000085:Acetates; D004791:Enzyme Inhibitors; D010940:Plant Proteins; D011799:Quinazolines; D047390:Casein Kinase II; D017346:Protein Serine-Threonine Kinases