title: Click chemistry mediated Eu-tagging: activity-based specific quantification and simultaneous activity evaluation of CYP3A4 using 153Eu species-unspecific isotope dilution inductively coupled plasma mass spectrometry.
abstract: P450 3A4 (CYP3A4) is one of the most important isoforms in the human cytochrome P450 superfamily. It was used as an example in this proof-of-concept study in order to demonstrate an activity-based labeling and then click chemistry (CC) mediated element-tagging strategy for simultaneously specific quantification and activity measurement of an enzyme using species-unspecific isotope dilution inductively coupled plasma mass spectrometry (SUID ICPMS). A dual functional hexynylated 17α-ethynylestradiol activity-based probe was synthesized for specifically labeling CYP3A4 and then CC-mediated Eu-tagging with an azido-DOTA-Eu complex for CYP3A4 quantification and activity measurement in human liver microsome and serum samples using (153)Eu SUID ICPMS. The LOD (3σ) of CYP3A4 reached 20.3 fmol when monitoring (151/153)Eu ICPMS signals, in addition to the merits of specificity and simultaneous activity measurement achieved. We believe that this activity-based CC-mediated element-tagging strategy will liberate more potential advantages of ICPMS in bioanalysis.
authors.affiliation: Department of Chemistry and the MOE Key Laboratory of Spectrochemical Analysis & Instrumentation and ‡The State Key Laboratory of Marine Environmental Science, Xiamen University , Xiamen 361005, China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Liang
authors.forename: Xiaowen
authors.initials: X
authors.lastname: Yan
authors.forename: Zhaoxin
authors.initials: Z
authors.lastname: Li
authors.forename: Limin
authors.initials: L
authors.lastname: Yang
authors.forename: Bo
authors.initials: B
authors.lastname: Zhang
authors.forename: Qiuquan
authors.initials: Q
authors.lastname: Wang
mesh_terms: D057930:Click Chemistry / Q000379:methods*; D051544:Cytochrome P-450 CYP3A / Q000737:chemistry* / Q000378:metabolism; D004958:Estradiol / Q000737:chemistry; D005063:Europium / Q000737:chemistry*; D006801:Humans; D007554:Isotopes; D011865:Radioisotope Dilution Technique; D011868:Radioisotopes; D013250:Steroid Hydroxylases / Q000737:chemistry / Q000378:metabolism
chemical_list: D007554:Isotopes; D011868:Radioisotopes; D005063:Europium; D004958:Estradiol; D013250:Steroid Hydroxylases; D051544:Cytochrome P-450 CYP3A; C022657:steroid hormone 6-beta-hydroxylase; C510163:CYP3A4 protein, human