A one-bead, one-stock solution approach to chemical genetics: part 1

Abstract

An efficient and accessible approach to split-pool, diversity-oriented synthesis using high-capacity macrobeads as individual microreactors has been developed. Each macrobead contains sufficient compound to generate a stock solution amenable to many biological assays, and reliable library encoding allows for rapid compound structure elucidation post-synthesis. This 'one-bead, one-stock solution' strategy is a central element of a technology platform aimed at advancing chemical genetics.

Domain-selective small-molecule inhibitor of histone deacetylase 6 (HDAC6)-mediated tubulin deacetylation

“…To verify TDAC-selective inhibitors, stock solutions of compounds active in the AcTubulin and͞or ITSA1ϩAcTubulin cytoblots but not the AcLysine cytoblot were retested in cells by using fluorescence microscopy. Beads corresponding to promising wells then were subjected to gas chromatographic decoding, leading to structure determination and subsequent resynthesis (15,16). One hydroxamic acid-containing compound, here named tubacin (1) (Fig.…”

“…14. Bead decoding and structure determination were as described (15,16). Tubacin and niltubacin were synthesized as described (15).…”

“…5D; refs. [13][14][15][16][17]. Two acetylation state-specific antibodies were exploited in cytoblot assays to distinguish between small molecules capable of inducing ␣-tubulin acetylation (AcTubulin) and histone acetylation (AcLysine).…”

Protein “fingerprinting” in complex mixtures with peptoid microarrays

“…These arrays are generally produced by first synthesizing a combinatorial library on a suitable bead resin, separating the beads into the wells of microtiter plates, and then releasing the compounds from the beads (4,5). The resulting solutions then are spotted robotically onto a chemically modified glass slide such that the library-derived molecule is attached covalently to the surface.…”

Chemical Biology/ Chemical Genetics/ Chemical Genomics: Importance of Chemical Library

“…If a molecular skeleton has multiple reactive sites with potential for orthogonal functionalization, then the technique of split-pool synthesis [62] [63] [64] can be used to harness the power of combinatorics and thereby generate all possible combinations of appendages efficiently [65]. The origins of DOS were combinatorial chemistry efforts began with a complexity-generating reaction to yield a single, complex molecular skeleton having several attachment points followed by a series of diversity-generating appending process (potentially in split-pool format) to attach all possible combinations of building blocks to this common skeleton.…”