毛细管电泳-激光诱导荧光检测用于多个胞内蛋白激酶的抑制剂筛选和选择性评价.pdfVIP

ZHANG Qianqian，eta1．：Inhibitorscreeningandselectivityassessmentagainst ses byca illary dectroph0re ．647 muhiplece arpr nkina p 第 7期 with laser—inducednnuuOorreesscceenncceedetection … · have become the second largestgroup ofthera— thecrudecelllysatesmaybeadesirablechoiceto peutictargets，after G—protein coupled receptors discoverthemulti—targeted inhibitors[17，18]． 『1，6]．Todate，ninesmallmolecularproteinki— Furthermore，quantifyingprotein kinaseactivities naseinhibitors(PKIs)havebeenapprovedbythe directly in complex celllysatesmaybehelpIfulto US Food and Drug Administration (FDA) for understand thebiologicalregulatory pathways of cancertreatm ent．andapproximately80inhibitors thePKIs． havebeeninvariousstagesofclinictrials[7]． Syntheticpeptidesw ith specific sequences are Despitesucha success，comparedto thenumber commonlyusedasthesubstratesforPKIscreen— of the clinically validated protein kinase—based inginbiochemicalassayformat[19—22]．Com— drugtargets，PKIshaving high potency and tea— pared to protein substrates，using peptide sub— sonableselectivitytowardtheirtargetkinasesare stratescan simplifyprotein kinaseassaym ethods quiterare[8]．Moreover，noneofthekinase—tar— due to the advantagessuch as ease ofstorage， geted drugsresponsible fortreating immunologi— handlingandprotocolmodification [5]．Several cal，neurological，metabolicandinfectiousdisea— substratepeptide-b·ased techniques such as fluo-· seshaveyetbeen proved by US FDA although rescence polarization and tim e--resolved fluores—- these diseases also involve dysregulated protein cence arewidely used forPKIscreening due to kinases[7，9]．Therefore，iti