Τίτλος:
Cloning and purification of protein kinase CK2 recombinant alpha and beta subunits from the Mediterranean fly Ceratitis capitata
Γλώσσες Τεκμηρίου:
Αγγλικά
Περίληψη:
The Mediterranean fruit fly Ceratitis capitata is an insect capable of wreaking extensive damage to a wide range of fruit crops. Protein kinase CK2 is a ubiquitous Ser/Thr kinase that is highly conserved among eukaryotes; it is a heterotetramer composed of two catalytic (α) and a dimer of regulatory (β) subunits. We present here the construction of the cDNA molecules of the CK2α and CK2β subunits from the medfly C. capitata by the 5′/3′ RACE and RT-PCR methods, respectively. CcCK2α catalytic subunit presents the characteristic and conserved features of a typical protein kinase, similar to the regulatory CcCK2β subunit, that also possess the conserved features of regulatory CK2β subunits, as revealed by comparison of their predicted amino acid sequences with other eukaryotic species. The recombinant CcCK2α and CcCK2β proteins were purified by affinity chromatography to homogeneity, after overexpression in Escherichia coli. CcCK2α is capable to utilize GTP and its activity and is inhibited by polyanions and stimulated by polycations in phosphorylation assays, using purified acidic ribosomal protein P1 as a substrate. © Springer Science+Business Media, LLC. 2011.
Συγγραφείς:
Kouyanou-Koutsoukou, S.
Baier, A.
Kolaitis, R.-M.
Maniatopoulou, E.
Thanopoulou, K.
Szyszka, R.
Περιοδικό:
Molecular and Cellular Biochemistry
Λέξεις-κλειδιά:
casein kinase 2 alpha; casein kinase 2 beta; casein kinase II; complementary DNA; guanosine triphosphate; polyanion; polycation; recombinant protein; ribosome protein; unclassified drug, affinity chromatography; amino acid sequence; article; controlled study; enzyme active site; enzyme activity; Escherichia coli; eukaryote; female; Mediterranean fruit fly; molecular cloning; nonhuman; nucleotide sequence; protein determination; protein expression; protein phosphorylation; protein purification; protein subunit; reverse transcription polymerase chain reaction, Amino Acid Sequence; Animals; Casein Kinase II; Catalytic Domain; Ceratitis capitata; Chromatography, Affinity; Cloning, Molecular; DNA, Complementary; Electrophoresis, Polyacrylamide Gel; Escherichia coli; Immobilized Proteins; Molecular Sequence Data; Protein Subunits; Recombinant Proteins; Sequence Alignment, Ceratitis capitata; Escherichia coli; Eukaryota; Hexapoda
DOI:
10.1007/s11010-011-0968-1
