TY  - JOUR
A1  - Meyners, Christian
A1  - Mertens, Monique
A1  - Wessig, Pablo
A1  - Meyer-Almes, Franz-Josef
T1  - A Fluorescence-Lifetime-Based Binding Assay for Class IIa Histone Deacetylases
JF  - Chemistry - a European journal
N2  - Class IIa histone deacetylases (HDACs) show extremely low enzymatic activity and no commonly accepted endogenous substrate is known today. Increasing evidence suggests that these enzymes exert their effect rather through molecular recognition of acetylated proteins and recruiting other proteins like HDAC3 to the desired target location. Accordingly, class IIa HDACs like bromodomains have been suggested to act as “Readers” of acetyl marks, whereas enzymatically active HDACs of class I or IIb are called “Erasers” to highlight their capability to remove acetyl groups from acetylated histones or other proteins. Small-molecule ligands of class IIa histone deacetylases (HDACs) have gained tremendous attention during the last decade and have been suggested as pharmaceutical targets in several indication areas such as cancer, Huntington's disease and muscular atrophy. Up to now, only enzyme activity assays with artificial chemically activated trifluoroacetylated substrates are in use for the identification and characterization of new active compounds against class IIa HDACs. Here, we describe the first binding assay for this class of HDAC enzymes that involves a simple mix-and-measure procedure and an extraordinarily robust fluorescence lifetime readout based on [1,3]dioxolo[4,5-f]benzodioxole-based ligand probes. The principle of the assay is generic and can also be transferred to class I HDAC8.
KW  - drug discovery
KW  - enzymes
KW  - fluorescent probes
KW  - high-throughput screening
KW  - hydrolases
Y1  - 2017
U6  - https://doi.org/10.1002/chem.201605140
SN  - 0947-6539
SN  - 1521-3765
VL  - 23
IS  - 13
SP  - 3107
EP  - 3116
PB  - Wiley-VCH
CY  - Weinheim
ER  - 
TY  - JOUR
A1  - Dai, Xiaolin
A1  - Mate, Diana M.
A1  - Glebe, Ulrich
A1  - Garakani, Tayebeh Mirzaei
A1  - Körner, Andrea
A1  - Schwaneberg, Ulrich
A1  - Böker, Alexander
T1  - Sortase-mediated ligation of purely artificial building blocks
JF  - Polymers
N2  - Sortase A (SrtA) from Staphylococcus aureus has been often used for ligating a protein with other natural or synthetic compounds in recent years. Here we show that SrtA-mediated ligation (SML) is universally applicable for the linkage of two purely artificial building blocks. Silica nanoparticles (NPs), poly(ethylene glycol) and poly(N-isopropyl acrylamide) are chosen as synthetic building blocks. As a proof of concept, NP-polymer, NP-NP, and polymer-polymer structures are formed by SrtA catalysis. Therefore, the building blocks are equipped with the recognition sequence needed for SrtA reaction-the conserved peptide LPETG-and a pentaglycine motif. The successful formation of the reaction products is shown by means of transmission electron microscopy (TEM), matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-ToF MS), and dynamic light scattering (DLS). The sortase catalyzed linkage of artificial building blocks sets the stage for the development of a new approach to link synthetic structures in cases where their synthesis by established chemical methods is complicated.
KW  - sortase-mediated ligation
KW  - enzymes
KW  - block copolymers
KW  - nanoparticles
Y1  - 2018
U6  - https://doi.org/10.3390/polym10020151
SN  - 2073-4360
VL  - 10
IS  - 2
PB  - MDPI
CY  - Basel
ER  - 
TY  - JOUR
A1  - Senf, Deborah
A1  - Ruprecht, Colin
A1  - Kishani, Saina
A1  - Matic, Aleksandar
A1  - Toriz, Guillermo
A1  - Gatenholm, Paul
A1  - Wagberg, Lars
A1  - Pfrengle, Fabian
T1  - Tailormade polysaccharides with defined branching patterns
BT  - Enzymatic polymerization of arabinoxylan oligosaccharides
JF  - Angewandte Chemie : a journal of the Gesellschaft Deutscher Chemiker ; International edition
N2  - The heterogeneous nature of non-cellulosic polysaccharides, such as arabinoxylan, makes it difficult to correlate molecular structure with macroscopic properties. To study the impact of specific structural features of the polysaccharides on crystallinity or affinity to other cell wall components, collections of polysaccharides with defined repeating units are required. Herein, a chemoenzymatic approach to artificial arabinoxylan polysaccharides with systematically altered branching patterns is described. The polysaccharides were obtained by glycosynthase-catalyzed polymerization of glycosyl fluorides derived from arabinoxylan oligosaccharides. X-ray diffraction and adsorption experiments on cellulosic surfaces revealed that the physicochemical properties of the synthetic polysaccharides strongly depend on the specific nature of their substitution patterns. The artificial polysaccharides allow structure-property relationship studies that are not accessible by other means.
KW  - carbohydrates
KW  - enzymes
KW  - glycosynthases
KW  - structure elucidation
KW  - synthetic methods
Y1  - 2018
U6  - https://doi.org/10.1002/anie.201806871
SN  - 1433-7851
SN  - 1521-3773
VL  - 57
IS  - 37
SP  - 11987
EP  - 11992
PB  - Wiley-VCH
CY  - Weinheim
ER  -