Johan M. Winne

V-ATPase-activity in the TGN/EE is required for exocytosis and recycling in Arabidopsis

In plants, vacuolar H+-ATPase (V-ATPase) activity acidifies both the trans-Golgi network/early endosome (TGN/EE) and the vacuole. This dual V-ATPase function has impeded our understanding of how the pH homeostasis within the plant TGN/EE controls exo- and endocytosis. Here, we show that the weak V-ATPase mutant deetiolated3 (det3) displayed a pH increase in the TGN/EE, but not in the vacuole, strongly impairing secretion and recycling of the brassinosteroid receptor and the cellulose synthase complexes to the plasma membrane, in contrast to mutants lacking tonoplast-localized V-ATPase activity only. The brassinosteroid insensitivity and the cellulose deficiency defects in det3 were tightly correlated with reduced Golgi and TGN/EE motility. Thus, our results provide strong evidence that acidification of the TGN/EE, but not of the vacuole, is indispensable for functional secretion and recycling in plants.

Scope and Mechanism of the (4+3) Cycloaddition Reaction of Furfuryl Cations†

The (4+3) cycloaddition reaction between a conjugated diene and an allylic cation constitutes a convenient and conceptually straightforward method to prepare seven-membered rings. 2] In recent years, investigations into this type of cycloaddition, including a number of elegant applications in total synthesis, have demonstrated its potential for development as a synthetic method that could approach the (4+2) cycloaddition (Diels–Alder) reactions in terms of selectivity and efficiency. Of particular interest are allylic cations bearing a cation-stabilizing substituent (Z group) at the 2-position [Eq. (1)] and stabilized allylic cations such as vinyloxocarbenium ions [Eq. (2)] or, typically, cationic reaction partners incorporating both of these features [Eq. (2), R = Z].

Chemical control of the viscoelastic properties of vinylogous urethane vitrimers

Vinylogous urethane based vitrimers are polymer networks that have the intrinsic property to undergo network rearrangements, stress relaxation and viscoelastic flow, mediated by rapid addition/elimination reactions of free chain end amines. Here we show that the covalent exchange kinetics significantly can be influenced by combination with various simple additives. As anticipated, the exchange reactions on network level can be further accelerated using either Brønsted or Lewis acid additives. Remarkably, however, a strong inhibitory effect is observed when a base is added to the polymer matrix. These effects have been mechanistically rationalized, guided by low-molecular weight kinetic model experiments. Thus, vitrimer elastomer materials can be rationally designed to display a wide range of viscoelastic properties.

Mitochondrial uncouplers inhibit clathrin-mediated endocytosis largely through cytoplasmic acidification

ATP production requires the establishment of an electrochemical proton gradient across the inner mitochondrial membrane. Mitochondrial uncouplers dissipate this proton gradient and disrupt numerous cellular processes, including vesicular trafficking, mainly through energy depletion. Here we show that Endosidin9 (ES9), a novel mitochondrial uncoupler, is a potent inhibitor of clathrin-mediated endocytosis (CME) in different systems and that ES9 induces inhibition of CME not because of its effect on cellular ATP, but rather due to its protonophore activity that leads to cytoplasm acidification. We show that the known tyrosine kinase inhibitor tyrphostinA23, which is routinely used to block CME, displays similar properties, thus questioning its use as a specific inhibitor of cargo recognition by the AP-2 adaptor complex via tyrosine motif-based endocytosis signals. Furthermore, we show that cytoplasm acidification dramatically affects the dynamics and recruitment of clathrin and associated adaptors, and leads to reduction of phosphatidylinositol 4,5-biphosphate from the plasma membrane.

Total Synthesis of (+/−)‐Frondosin B and (+/−)‐5‐epi‐Liphagal by Using a Concise (4+3) Cycloaddition Approach

A recently developed (4+3) cycloaddition between dienes and furfuryl alcohols, as precursors of oxyallyl-type cations, has been used as a key step in the racemic syntheses of two natural products: frondosin B and liphagal. This work demonstrates the synthetic potential of this cycloaddition reaction, and offers a short synthetic route to an interesting family of natural products. A full account of these synthetic studies is presented, further illustrating the mechanism, scope, and limitations of this straightforward synthetic method for seven-membered rings.

Possibility of [1,5] sigmatropic shifts in bicyclo[4.2.0]octa-2,4-dienes.

The thermal equilibration of the methyl esters of endiandric acids D and E was subject to a computational study. An electrocyclic pathway via an electrocyclic ring opening followed by a ring flip and a subsequent electrocyclization proposed by Nicolaou [ Nicolaou , K. C. ; Chen , J. S. Chem. Soc. Rev. 2009 , 38 , 2993 ], was computationally explored. The free-energy barrier for this electrocyclic route was shown to be very close to the bicyclo[4.2.0]octa-2,4-diene reported by Huisgen [ Huisgen , R. ; Boche , G. ; Dahmen , A. ; Hechtl , W. Tetrahedron Lett. 1968 , 5215 ]. Furthermore, the possibility of a [1,5] sigmatropic alkyl group shift of bicyclo[4.2.0]octa-2,4-diene systems at high temperatures was explored in a combined computational and experimental study. Calculated reaction barriers for an open-shell singlet biradical-mediated stepwise [1,5] sigmatropic alkyl group shift were shown to be comparable with the reaction barriers for the bicyclo[4.1.0]hepta-2,4-diene (norcaradiene) walk rearrangement. However, the stepwise sigmatropic pathway is suggested to only be feasible for appropriately substituted compounds. Experiments conducted on a deuterated analogous diol derivative confirmed the calculated (large) differences in barriers between electrocyclic and sigmatropic pathways.

Polydimethylsiloxane quenchable vitrimers

An elastomeric vitrimer material was synthesized via the crosslinking of a polydimethylsiloxane bearing pendant amino functions with a bis-vinylogous urethane crosslinker. The elastic properties of this material could be tuned across a wide range of moduli by varying crosslink density. Reshaping and recyclability were observed at 100 °C. Further investigation of the dynamic character revealed double relaxations, which could be supressed by masking free amino groups. Finally, these studies led to the development of a single-pot, solution-castable formulation of a non-dynamic permanent elastomer. These results open a way for the design of quenchable vitrimers, that is to say crosslinked materials which after shaping or recycling exhibit conventional elastomer properties.

Nonenzymic polycyclisation of analogues of oxidosqualene with a preformed C-ring

Some nonenzymic epoxide-initiated polyolefin cyclization are reported. The presented molecules are partially constrained analogues of (3S)-oxidosqualene, the natural substrate to many important cyclase enzymes. These model compounds feature a preformed C-ring with built-in stereochemical information. The experimental results allow for an instructive comparison with the enzymic processes, particularly those of the cyclases in steroid biosynthesis (i.e. lanosterol synthase).

Covalent Fluorination Strategies for the Surface Modification of Polydienes

Nonreactive additives are widely applied to enhance polymer properties but can leach out of the material over time. In this work, two essentially different fluorinated additives bearing a triazolinedione moiety are synthesized and grafted on several polydiene backbones (acrylonitrile-butadiene-styrene, styrene-butadiene, and styrene-isoprene-styrene (SIS) copolymers), either by dip-coating or by reaction in solution. The resulting polymers are analyzed by contact angle goniometry, size exclusion chromatography, and NMR, infrared, and X-ray photoelectron spectroscopy. Independent of the modification procedure, the fluorophilic perfluoroalkyl additive is found at the material surface, thereby yielding a more hydrophobic surface. For SIS thermoplastic elastomers, for example, contact angles up to 125° can be obtained.

(5,6-Dihydro-1,4-dithiin-2-yl)methanol as a Versatile Allyl-Cation Equivalent in (3+2) Cycloaddition Reactions.

The title heterocyclic alcohol readily generates a sulfur-substituted allylic cation upon simple treatment with a protic acid, thus facilitating a synthetically useful stepwise (3+2) cycloaddition reaction pathway with a range of conjugated-olefin-type substrates. The introduction of an allyl fragment in this way provided rapid access to a variety of cyclopentanoid scaffolds. The cyclic nature of the cation precursor alcohol was shown to be instrumental for efficient cycloaddition reactions to take place, thus indicating an attractive strategy for controlling the reactivity of heteroatom-substituted allyl cations. The formal cycloaddition reaction is highly regio- and stereoselective and was also used for a short total synthesis of the natural product cuparene in racemic form through a cycloaddition-hydrodesulfurization sequence.