Optimization of Protein-Protein Interaction Measurements for Drug Discovery Using AFM Force Spectroscopy

Increasingly targeted in drug discovery, protein-protein interactions challenge current high throughput screening technologies in the pharmaceutical industry. Developing an effective and efficient method for screening small molecules or compounds is critical to accelerate the discovery of ligands for enzymes, receptors and other pharmaceutical targets. Here, we report developments of methods to increase the signal-to-noise ratio (SNR) for screening protein-protein interactions using atomic force microscopy (AFM) force spectroscopy. We have demonstrated the effectiveness of these developments on detecting the binding process between focal adhesion kinases (FAK) with protein kinase B (Akt1), which is a target for potential cancer drugs. These developments include optimized probe and substrate functionalization processes and redesigned probe-substrate contact regimes. Furthermore, a statistical-based data processing method was developed to enhance the contrast of the experimental data. Collectively, these results demonstrate the potential of the AFM force spectroscopy in automating drug screening with high throughput

More microbial manipulation and plant defense than soil fertility for biochar in food production: A field experiment of replanted ginseng with different biochars

The role of biochar–microbe interaction in plant rhizosphere mediating soilborne disease suppression has been poorly understood for plant health in field conditions. Chinese ginseng ( Panax ginseng C. A. Meyer) is widely cultivated in Alfisols across Northeast China, being often stressed severely by pathogenic diseases. In this study, the topsoil of a continuously cropped ginseng farm was amended at 20 t ha − 1, respectively, with manure biochar (PB), wood biochar (WB), and maize residue biochar (MB) in comparison to conventional manure compost (MC). Post-amendment changes in edaphic properties of bulk topsoil and the rhizosphere, in root growth and quality, and disease incidence were examined with field observations and physicochemical, molecular, and biochemical assays. In the 3 years following the amendment, the increases over MC in root biomass were parallel to the overall fertility improvement, being greater with MB and WB than with PB. Differently, the survival rate of ginseng plants increased insignificantly with PB but significantly with WB (14%) and MB (21%), while ginseng root quality was unchanged with WB but improved with PB (32%) and MB (56%). For the rhizosphere at harvest following 3 years of growing, the total content of phenolic acids from root exudate decreased by 56, 35, and 45% with PB, WB, and MB, respectively, over MC. For the rhizosphere microbiome, total fungal and bacterial abundance both was unchanged under WB but significantly increased under MB (by 200 and 38%), respectively, over MC. At the phyla level, abundances of arbuscular mycorrhizal and Bryobacter as potentially beneficial microbes were elevated while those of Fusarium and Ilyonectria as potentially pathogenic microbes were reduced, with WB and MB over MC. Moreover, rhizosphere fungal network complexity was enhanced insignificantly under PB but significantly under WB moderately and MB greatly, over MC. Overall, maize biochar exerted a great impact rather on rhizosphere microbial community composition and networking of functional groups, particularly fungi, and thus plant defense than on soil fertility and root growth

More microbial manipulation and plant defense than soil fertility for biochar in food production: A field experiment of replanted ginseng with different biochars

The role of biochar–microbe interaction in plant rhizosphere mediating soil-borne disease suppression has been poorly understood for plant health in field conditions. Chinese ginseng (Panax ginseng C. A. Meyer) is widely cultivated in Alfisols across Northeast China, being often stressed severely by pathogenic diseases. In this study, the topsoil of a continuously cropped ginseng farm was amended at 20 t ha–1, respectively, with manure biochar (PB), wood biochar (WB), and maize residue biochar (MB) in comparison to conventional manure compost (MC). Post-amendment changes in edaphic properties of bulk topsoil and the rhizosphere, in root growth and quality, and disease incidence were examined with field observations and physicochemical, molecular, and biochemical assays. In the 3 years following the amendment, the increases over MC in root biomass were parallel to the overall fertility improvement, being greater with MB and WB than with PB. Differently, the survival rate of ginseng plants increased insignificantly with PB but significantly with WB (14%) and MB (21%), while ginseng root quality was unchanged with WB but improved with PB (32%) and MB (56%). For the rhizosphere at harvest following 3 years of growing, the total content of phenolic acids from root exudate decreased by 56, 35, and 45% with PB, WB, and MB, respectively, over MC. For the rhizosphere microbiome, total fungal and bacterial abundance both was unchanged under WB but significantly increased under MB (by 200 and 38%), respectively, over MC. At the phyla level, abundances of arbuscular mycorrhizal and Bryobacter as potentially beneficial microbes were elevated while those of Fusarium and Ilyonectria as potentially pathogenic microbes were reduced, with WB and MB over MC. Moreover, rhizosphere fungal network complexity was enhanced insignificantly under PB but significantly under WB moderately and MB greatly, over MC. Overall, maize biochar exerted a great impact rather on rhizosphere microbial community composition and networking of functional groups, particularly fungi, and thus plant defense than on soil fertility and root growth