Purification and kinase assay of PKN.
PKN is a serine/threonine protein kinase, which has a catalytic domain highly homologous to that of protein kinase C (PKC) in the carboxyl-terminal region and three repeats of the antiparallel coiled coil (ACC) domain in the amino-terminal region. Mammalian PKN has three isoforms each derived from different genes, PKN1 (PKNalpha/PRK1/PAK1), PKN2 (PRK2/PAK2/PKNgamma), and PKN3 (PKNbeta). PKN isoforms show different enzymatic properties and tissue distributions and have been implicated in various distinct cellular processes (reviewed in Mukai [2003]). This chapter discusses methods to prepare purified enzymes and to assay substrate phosphorylation activities.
Mukai
Hideyuki
H
Biosignal Research Center, Kobe University, Japan.
Ono
Yoshitaka
Y
D000818:Animals; D001921:Brain / Q000201:enzymology; D002417:Cattle; D003600:Cytosol / Q000201:enzymology; D004789:Enzyme Activation; D006801:Humans; D007527:Isoenzymes / Q000378:metabolism; D008297:Male; D011493:Protein Kinase C; D017346:Protein Serine-Threonine Kinases / Q000302:isolation & purification* / Q000378:metabolism*; D011505:Protein-Tyrosine Kinases / Q000302:isolation & purification* / Q000378:metabolism*; D051381:Rats; D011994:Recombinant Proteins / Q000302:isolation & purification; D018411:Spodoptera; D013737:Testis / Q000201:enzymology
D007527:Isoenzymes; D011994:Recombinant Proteins; C061570:protein kinase N; D011505:Protein-Tyrosine Kinases; D017346:Protein Serine-Threonine Kinases; D011493:Protein Kinase C