Dataset Information

Simple , Enantiocontrolled Azetidine Library Synthesis via Strain-Release Functionalization of 1-Azabicyclobutanes

ABSTRACT: Herein we describe a new enantioselective route for the synthesis of all stereoisomers of 2,3-disubstituted azetidines. This approach uses simple strain release functionalization of preformed 1-azabicyclobutanes (ABBs), which enables installation of a wide array of nucleophiles at the C3-position. Chemical proteomic profiling of stereoisomeric sets of cysteine-reactive acrylamides derived from the new azetidine library reveals ligandable protein sites distinct from those of azetidine probes synthesized using C–H arylation.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Suspension Culture, Lymphocyte Of B Lineage

SUBMITTER: Hayden Sharma

LAB HEAD: Dr. Benjamin F. Cravatt

PROVIDER: PXD055926 | Pride | 2026-06-29

REPOSITORIES: Pride

ACCESS DATA

Dataset's files

Source:

			Action	DRS
	20240510_has-ii-259_01.raw	Raw
	20240510_has-ii-259_02.raw	Raw
	20240510_has-ii-259_03.raw	Raw
	20240510_has-ii-259_04.raw	Raw
	20240510_has-ii-259_05.raw	Raw

Items per page:

1 - 5 of 46

Publications

Enantiocontrolled Azetidine Library Synthesis via Strain-Release Functionalization of 1-Azabicyclobutanes.

Bielecki Michael M Nassir Molhm M Sharma Hayden A HA Truax Nathanyal J NJ Raheja Nicholas N Thompson Ty M TM El-Hayek Ewing Tamara T Melillo Bruno B Cravatt Benjamin F BF Baran Phil S PS

Journal of the American Chemical Society 20250611 25

A simple, modular, and programmable approach to access complex stereopure azetidines through strain-release functionalization is disclosed. The synthetic methods developed enable the parallel synthesis of stereodefined azetidines that would be otherwise laborious to produce. Given the privileged nature of these structures, a set of stereoprobes for use in activity-based protein profiling was prepared and evaluated, revealing proteins in human cancer cells, which were liganded with clear stereo- ...[more]

PMID: 40501015

Similar Datasets

Project description:The dynamic protein distribution within and across the plasma membrane is pivotal in regulating cell communications. Yet, methods facilitating rapid labeling, applicable across diverse cell types, and amenable to multiplexed live imaging are scarce. We discovered that live mammalian cell surface proteins can be labeled using N-hydroxysuccinimide (NHS)-ester-based amine crosslinking of fluorescent dyes. Using mouse DC2.4 dendritic cells, human Jurkat T cells, and mouse primary lymphoma T cells as model systems, we revealed the membrane topology and membrane-associated cell communications previously challenging to capture by fluorescence microscopy techniques. In DC2.4 cells, we observed transient membrane protein accumulation at cell-cell contacts and their bidirectional migration patterns guided by membrane fibers. We further visualized the involvement of membrane fibers in the transfer of membrane-derived particles and ovalbumin, as well as caveolin-dependent endocytosis and subsequent transport of membrane labels in late endosomes and lysosomes. In addition, we demonstrate caveolin-1 phosphorylation-dependent insulin receptor endocytosis in HEK 293T cells. Volumetric structured illumination microscopy revealed changes in the membrane topology of Jurkat T cells in response to the activating surface, generation of membrane-derived microvesicles, and phagocytosis of CD3/CD28 Dynabeads on the glass surface. Finally, we demonstrate that the membrane label remains stable in the in vivo environment and evidence of intercellular membrane protein transfer from splenocytes from the donor T cell lymphoma SNF5 mouse to the healthy splenocytes in the C57/BL6 mouse 24 and 48-h after cell transfer. The ultrahigh membrane labeling density, coupled with high multiplexity and rapid labeling, enables innovative fluorescence microscopy applications to dissect membrane and membrane-derived structures in intercellular and intracellular communication both in vitro and in vivo.

Project description:Dialkyldiazirines have emerged as a photoaffinity probe of choice for interactome mapping in live cell experiments. Upon irradiation, ‘linear’ dialkyldiazirines produce dialkylcarbenes, which are susceptible to both intramolecular reactions and unimolecular elimination processes, as well as diazoalkanes, which also participate in intermolecular labeling. Cyclobutylidene has a nonclassical bonding structure and is stable enough to be captured in bimolecular reactions. Cyclobutanediazirines have more recently been studied as photoaffinity probes based on cyclobutylidene, but the mechanism, especially with respect to the role of putative diazo intermediates, was not fully understood. Here, we show that photolysis (365 nm) of cyclobutanediazirines can produce cyclobutylidene intermediates as evidenced by formation of their expected bimolecular and unimolecular products, including methylenecyclopropane derivatives. Unlike linear diazirines, cyclobutanediazirine photolysis in the presence of tetramethylethylene produces a [2+1] cycloaddition adduct. By contrast, linear diazirines produce diazo compounds upon low temperature photolysis in THF, whereas diazo compounds are not detected in similar photolyses of cyclobutanediazirines. Diazocyclobutane, prepared by independent synthesis, is labile, reactive toward water, and capable of protein alkylation. The rate of diazocyclobutane decomposition is not affected by 365 nm light, suggesting that the photochemical conversion of diazocyclobutane to cyclobutylidene is not an important pathway. Finally, chemical proteomic studies revealed that a likely consequence of this primary conversion to a highly reactive carbene is a marked decrease in labeling by cyclobutanediazirine-based probes relative to linear diazirine counterparts both at the individual protein and proteome-wide levels. Collectively, these observations are consistent with a mechanistic picture for cyclobutanediazirine photolysis that involves carbene chemistry with minimal formation of diazo intermediates, and contrasts with the photolyses of linear diazirines where alkylation by diazo intermediates plays a more significant role.