酪氨酸磷酸化多肽的化学选择性富集.pdfVIP

· 870 · 色 谱 第 29卷 m ethods，such as immobilized m etalion affi nity been used ofr serine and threonine phosphopep— chromatography(IMAC)[3—6]andtheuseof tideenrichment [12]．Underbasicconditions， metaloxideparticles[7—9]，havebeendevel— phosphothreonine／phosphoserine first undergoes oped to selectively enrich phosphopeptides from 一 elimination (Fig．1)andfollowedbyMichael complex m ixtures．How ever，none ofthe above addition in the presence ofa nucleophile which ， approaches can targetthe certain type ofphos— allowsaffinitytags (e．g．，biotin)tobeintro— phopeptides． Anti—phosphOtyrosine antibody ducedon form erly phosphorylated sites．Finally， based m ethod has been recognized as the most functionalized beads can be used to capture the effective m ethod for the enrichm ent of tyrosine form erly form ed serine／threonine phosphopep— phosphorylatedproteinsandpeptides[10]．How— tides．Herein，instead ofthe enrichm entof ser— ever，the high cost of antibody—based method ine／threoninephosphopeptides， 一elim inationwas decreasesitsapplicabilityforroutineuse，and its used to remove the phospho—groups on serine／ repr0ducibility cannot be wellcontrolled． More threonineresidues[13]．Then，ahighlyefficient im portantly，tyrosine antibodies for imm unopre· affinity approach was used to enrich tyrosine cipitation are biased to certain motifs． It may phosphopeptidesleftin the sample．The 一elim i— need to combine severalantibodies to getmore nation reactionw asoptim ized ofr complete reac— com prehensive enrichment． Therefore， there is