{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,15]],"date-time":"2026-09-15T20:50:51Z","timestamp":1789505451589,"version":"build-2803163510"},"reference-count":35,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2015,2,16]],"date-time":"2015-02-16T00:00:00Z","timestamp":1424044800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["chemistry-europe.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["ChemBioChem"],"published-print":{"date-parts":[[2015,3,23]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>We report the evaluation of two alternative chemical dimerizer approaches aimed at increasing the sensitivity of MASPIT, a three\u2010hybrid system that enables small\u2010molecule target protein profiling in intact human cells. To circumvent the potential limitations related to the binding of methotrexate (MTX) to endogenous human dihydrofolate reductase (DHFR), we explored trimethoprim (TMP) as an alternative prokaryote\u2010specific DHFR ligand. MASPIT evaluation of TMP fusion compounds with tamoxifen, reversine, and simvastatin as model baits, resulted in dose\u2013response curves shifted towards lower EC<jats:sub>50<\/jats:sub> values than those of their MTX congeners. Furthermore, a scalable azido\u2010TMP reagent was synthesized that displayed a similar improvement in sensitivity, possibly owing to increased membrane permeability relative to the MTX anchor. Applying the SNAP\u2010tag approach to introduce a covalent bond into the system, on the other hand, produced an inferior readout than in the MTX\u2010 or TMP\u2010tag based assay.<\/jats:p>","DOI":"10.1002\/cbic.201402702","type":"journal-article","created":{"date-parts":[[2015,2,16]],"date-time":"2015-02-16T15:47:40Z","timestamp":1424101660000},"page":"834-843","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Alternative Reagents for Methotrexate as Immobilizing Anchor Moieties in the Optimization of MASPIT: Synthesis and Biological Evaluation"],"prefix":"10.1002","volume":"16","author":[{"given":"Dries J. H.","family":"De Clercq","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Martijn D. 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