蛋白質(zhì)及其標(biāo)記ppt課件-資料下載頁(yè)
2025-01-19 01:39本頁(yè)面
【正文】 the same azide ponent as the azidephosphine (Staudinger) chemistry, it requires special coppercontaining reaction buffers that have damaging effects on cellular ponents. Features Benefits ? Chemoselective reactivity ?Virtually no side reactions with other functional groups ?Detect and enrich without contamination ?Accurate and representative biological observations ? Compatible with bioorthogonal metabolic labeling ?Terminal azido functional groups not found in living systems ?Azido derivatives of biological molecules are well tolerated ? High reaction efficiency ?Close to 100% reaction efficiency in chemical model systems ? Ease of use ?Tworeagent system is easier to use and troubleshoot than alternative coppercatalyzed click chemistry (5 different reagents) ? Biologicalsample patibility ?Reactions occur in aqueous solutions at physiological pH with no known side reactions with buffers/detergents ?No risk for toxicity and protein oxidation because of the absence of Cu(I) ? Allows probe switching ? Azidolabeled molecules can be derivatized with wide variety of phosphine probes AzidePhosphine (Staudinger Ligation) SiteSpecific Protein Labeling and Protein Interaction Detection with Biotin Ligase Labeling cell surface proteins by acceptor peptide (AP) tagging with ketone 1 by BirA, followed by ligation of hydrazine derivatives to ketone 其他特殊標(biāo)記方法： 酶催化的特異性標(biāo)記 ALICE Y TING, MIT Nature Methods 2022, 2, 99104. PRIME ： PRobe Incorporation Mediated by Enzymes an engineered fluorophore ligase capable of recognizing a 7hydroxycoumarin substrate and catalyzing its covalent conjugation to a transposable 13amino acid peptide called LAP (LplA Acceptor Peptide) Uttamapinant, C. et al. Proc. Natl. Acad. Sci. USA 2022, 107, 10914 SnapTag The SNAPtag is an engineered mutant of the human repair protein O6alkylguanineDNA alkyltransferase (hAGT) which reacts with O6benzylguanine (BG) or O6benzyl4chloropyrimidine (CP) substrates modified with a synthetic label (., fluorophores, quantum dots, affinity ligands, and gold nanoparticles).
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