INT Home | Legals | Sitemap | INT-ra Net | KIT

Ligand-lipid and ligand-core affinity control the interaction of gold nanoparticles with artificial lipid bilayers and cell membranes

gold nanoparticle
Ligand-nanoparticle interactions control nanoparticle embedding in lipid membranes

Interactions between nanoparticles (NPs) and biomembranes depend on the physicochemical properties of the NPs, such as size and surface charge. Here we report on the size-dependent interaction of gold nanoparticles (AuNPs), stabilized with ligands differing in charge, i.e. sodium 3-(diphenylphosphino)benzene sulfonate (TPPMS) and sodium 3,3′,3″-triphenylphosphine sulfonate (TPPTS), respectively, with artificial membranes (black lipid membranes; BLMs) and HeLa cells. The TPPTS-stabilized AuNPs affect BLMs at lower size than TPPMS-stabilized ones. On HeLa cells we found decreasing cytotoxicity with increasing particle size, however, with an overall lower cytotoxicity for TPPTS-stabilized AuNPs. We attribute size-dependent BLM properties as well as reduced cytotoxicity of TPPTS-stabilized AuNPs to weaker shielding of the AuNP core when stabilized with TPPTS. We hypothesize that the partially unshielded hydrophobic gold core can embed into the hydrophobic membrane interior. Thereby we demonstrate that ligand-dependent cytotoxicity of NP can occur even when the NPs are not translocated through the membrane.

Janine Broda, Julia Setzler, Annika Leifert, Julia Steitz, Roland Benz, Ulrich Simon, Wolfgang Wenzel

Structure based design of protein linkers for zinc finger nuclease

ZFP design approach
ZFP design approach

Zinc finger nucleases are a promising tool to edit DNA in many biological applications, in particular for gene knockout. Despite many efforts the number of genes that can be effectively targeted with ZFNs remains severely limited, as available constructs cannot address arbitrary gene sequences. Here, we develop a novel concept to significantly enhance the number of DNA sequences that can be targeted by ZFN. Using an efficient computational model, we provide an extensive library of possible linker molecules between individual zinc finger motifs in the construct that can skip up to 10 base pairs between adjacent zinc finger recognition sites in the DNA sequence, which increases the number of genes that can be efficiently targeted by more than an order of magnitude.

Anand, P., Schug, A., and Wenzel, W.

Folding and Self-Assembly of the TatA Translocation Pore Based on a Charge Zipper Mechanism

Charge zippers as a new concept for folding and assembly of membrane proteins

We propose a concept for the folding and self-assembly of the pore-forming TatA complex from the Twin-arginine translocase and of other membrane proteins based on electrostatic “charge zippers.” Each subunit of TatA consists of a transmembrane segment, an amphiphilic helix (APH), and a C-terminal densely charged region (DCR). The sequence of charges in the DCR is complementary to the charge pattern on the APH, suggesting that the protein can be “zipped up” by a ladder of seven salt bridges. The length of the resulting hairpin matches the lipid bilayer thickness, hence a transmembrane pore could self-assemble via intra- and intermolecular salt bridges. The steric feasibility was rationalized by molecular dynamics simulations, and experimental evidence was obtained by monitoring the monomer-oligomer equilibrium of specific charge mutants. Similar “charge zippers” are proposed for other membrane-associated proteins, e.g., the biofilm-inducing peptide TisB, the human antimicrobial peptide dermcidin, and the pestiviral ERNS protein.

Grage S. L., Walther T. H., Klein M. J. , Wolf M. , Vargu A. , Ruggerone P. , Wenzel W., Ulrich A.S.

Cell 152, 316-326 (2013)

Differential hERG ion channel activity of ultrasmall gold nanoparticles

Molecular simulation of nanoparticle docking to the intracellular hERG channel

Understanding the mechanism of toxicity of nanomaterials remains a challenge with respect to both mechanisms involved and product regulation. Here we show toxicity of ultrasmall gold nanoparticles (AuNPs). Depending on the ligand chemistry, 1.4-nm-diameter AuNPs failed electrophysiology-based safety testing using human embryonic kidney cell line 293 cells expressing human ether-á-go-go-Related gene (hERG), a Food and Drug Administration-established drug safety test. In patch-clamp experiments, phosphine-stabilized AuNPs irreversibly blocked hERG channels, whereas thiol-stabilized AuNPs of similar size had no effect in vitro, and neither particle blocked the channel in vivo. We conclude that safety regulations may need to be reevaluated and adapted to reflect the fact that the binding modality of surface functional groups becomes a relevant parameter for the design of nanoscale bioactive compounds.

Leifert A, Pan Y, Kinkeldey A, Schiefer F, Setzler J, Scheel O, Lichtenbeld H, Schmid G, Wenzel W, Jahnen-Dechent W, Simon U.

PNAS 110, 8004-8009

7-Alkyl-3-benzylcoumarins: A Versatile Scaffold for the Development of Potent and Selective Cannabinoid Receptor Agonists and Antagonists


A series of 7-alkyl-3-benzylcoumarins was designed, synthesized, and tested at cannabinoid CB(1) and CB(2) receptors in radioligand binding and cAMP accumulation studies. 7-Alkyl-3-benzylcoumarins were found to constitute a versatile scaffold for obtaining potent CB receptor ligands with high potency at either CB(1) or CB(2) and a broad spectrum of efficacies. Fine-tuning of compound properties was achieved by small modifications of the substitution pattern. The most potent compounds of the present series include 5-methoxy-3-(2-methylbenzyl)-7-pentyl-2H-chromen-2-one (19a, PSB-SB-1201), a selective CB(1)antagonist (K(i) CB(1) 0.022 μM), 5-methoxy-3-(2-methoxybenzyl)-7-pentyl-2H-chromen-2-one (21a, PSB-SB-1202), a dual CB(1)/CB(2)agonist (CB(1)K(i) 0.032 μM, EC(50) 0.056 μM; CB(2)K(i) 0.049 μM, EC(50) 0.014 μM), 5-hydroxy-3-(2-hydroxybenzyl)-7-(2-methyloct-2-yl)-2H-chromen-2-one (25b, PSB-SB-1203), a dual CB(1)/CB(2) ligand that blocks CB(1) but activates CB(2) receptors (CB(1)K(i) 0.244 μM; CB(2)K(i) 0.210 μM, EC(50) 0.054 μM), and 7-(1-butylcyclopentyl)-5-hydroxy-3-(2-hydroxybenzyl)-2H-chromen-2-one (27b, PSB-SB-1204), a selective CB(2) receptor agonist (CB(1)K(i) 1.59 μM; CB(2)K(i) 0.068 μM, EC(50) 0.048 μM).

Rempel V., Volz N., Hinz S., Karcz T., Meliciani I., Nieger M., Wenzel W., Brase S., Muller C. E.

Journal of Medicinal Chemistry 55, 7967-7977 (2012)

In Silico Discovery of a Compound with Nanomolar Affinity to Antithrombin Causing Partial Activation and Increased Heparin Affinity

heparin tmi
Isothermal titration calorimetry confirmed a TMI higher than the heparin affinity

The medical and socioeconomic relevance of thromboembolic disorders promotes an ongoing effort to develop new anticoagulants. Heparin is widely used as activator of antithrombin but incurs side effects. We screened a large database in silico to find alternative molecules and predicted d-myo-inositol 3,4,5,6-tetrakisphosphate (TMI) to strongly interact with antithrombin. Isothermal titration calorimetry confirmed a TMI affinity of 45 nM, higher than the heparin affinity (273 nM). Functional studies, fluorescence analysis, and citrullination experiments revealed that TMI induced a partial activation of antithrombin that facilitated the interaction with heparin and low affinity heparins. TMI improved antithrombin inhibitory function of plasma from homozygous patients with antithrombin deficiency with a heparin binding defect and also in a model with endothelial cells. Our in silico screen identified a new, non-polysaccharide scaffold able to interact with the heparin binding domain of antithrombin. The functional consequences of this interaction were experimentally characterized and suggest potential anticoagulant therapeutic applications.

Navarro-Fernandez J., Perez-Sanchez H., Martinez-Martinez I., Meliciani I., Guerrero J. A., Vicente V., Corral J., Wenzel W.Journal of

Medicinal Chemistry 55, 6403-6412 (2012)

Grid Computing

3min Introduction into Protein Folding Simulations for a General Public

The video introduces protein folding simulations for a general public audience ("high-school level").