Femtosecond Structural Dynamics of Proteins

Proteins are the workhorses of living cells, providing essential functions such as structural support, signal transduction, enzymatic catalysis, transport and storage of small ligands. Atomic-resolution structures obtained with conventional X-ray crystallography show proteins essentially as static. In reality, however, proteins move and their motion is crucial for functioning. Although the structure and dynamics of proteins are intimately related, they are not equally well understood. A very large number of protein structures have been determined, but only a few studies have been able to monitor experimentally the dynamics of proteins in real time. In the last two decades, the availability of short (~100 ps) and intense (~10⁹–10¹⁰ photons) X-ray pulses produced by third-generation synchrotrons have allowed the implementation of structural methods like time-resolved X-ray crystallography and time-resolved X-ray solution scattering that have allowed us to monitor protein motion in the nanosecond-to-millisecond timescale [1 K. Moffat, Chem. Rev. 101, 1569–1582 (2001).[Crossref], [PubMed], [Web of Science ®] , [Google Scholar]–4 J. G. Kim, Acc. Chem. Res. 48, 2200–2208 (2015).[Crossref], [PubMed], [Web of Science ®] , [Google Scholar]]. Time-resolved X-ray crystallography has been used to monitor processes such as the migration of a ligand from the protein active site to the surrounding solvent [5 V. Srajer, Science 274, 1726–1729 (1996).[Crossref], [PubMed], [Web of Science ®] , [Google Scholar]–7 D. Bourgeois, Proc. Natl. Acad. Sci. U. S. A. 100, 8704–8709 (2003).[Crossref], [PubMed], [Web of Science ®] , [Google Scholar]] or tertiary structural changes associated with allosteric transitions [8 J. E. Knapp, Proc. Natl. Acad. Sci. 103, 7649–7654 (2006).[Crossref], [PubMed], [Web of Science ®] , [Google Scholar], 9 J. G. Kim, Struct. Dyn. 3, 023610 (2016). [Google Scholar]]. On the other hand, time-resolved X-ray scattering in the so-called wide-angle X-ray scattering (WAXS) region [10 M. Cammarata, Nat. Methods 5, 881–886 (2008).[Crossref], [PubMed], [Web of Science ®] , [Google Scholar]] has been used to track conformational changes corresponding to large-amplitude protein motions such as the quaternary R-T transition of human hemoglobin [11 M. Cammarata, J. Mol. Biol. 400, 951–962 (2010).[Crossref], [PubMed], [Web of Science ®] , [Google Scholar]–13 A. Spilotros, Soft Matter 8, 6434–6437 (2012).[Crossref], [Web of Science ®] , [Google Scholar]], the relative motion of bacteriorhodopsin α-helices following retinal isomerization [14 M. Andersson, Structure 17, 1265–1275 (2009).[Crossref], [PubMed], [Web of Science ®] , [Google Scholar]], or the open-to-close transition in bacterial phytochromes [15 H. Takala, Nature 509, 245–8 (2014).[Crossref], [PubMed], [Web of Science ®] , [Google Scholar]].     

A Method for Detecting Circulating Tumor Cells Based on the Measurement of Single‐Cell Metabolism in Droplet‐Based Microfluidics

The number of circulating tumor cells (CTCs) in blood is strongly correlated with the progress of metastatic cancer. Current methods to detect CTCs are based on immunostaining or discrimination of physical properties. Herein, a label‐free method is presented exploiting the abnormal metabolic behavior of cancer cells. A single‐cell analysis technique is used to measure the secretion of acid from individual living tumor cells compartmentalized in microfluidically prepared, monodisperse, picoliter (pL) droplets. As few as 10 tumor cells can be detected in a background of 200 000 white blood cells and proof‐of‐concept data is shown on the detection of CTCs in the blood of metastatic patients.     

Synergistic Effect of Multi-Walled Carbon Nanotubes and Ladder-Type Conjugated Polymers on the Performance of N-Type Organic Electrochemical Transistors

Organic electrochemical transistors (OECTs) have the potential to revolutionize the field of organic bioelectronics. To date, most of the reported OECTs include p-type (semi-)conducting polymers as the channel material, while n-type OECTs are yet at an early stage of development, with the best performing electron-transporting materials still suffering from low transconductance, low electron mobility, and slow response time. Here, the high electrical conductivity of multi-walled carbon nanotubes (MWCNTs) and the large volumetric capacitance of the ladder-type π-conjugated redox polymer poly(benzimidazobenzophenanthroline) (BBL) are leveraged to develop n-type OECTs with record-high performance. It is demonstrated that the use of MWCNTs enhances the electron mobility by more than one order of magnitude, yielding fast transistor transient response (down to 15 ms) and high μC* (electron mobility × volumetric capacitance) of about 1 F cm^?1 V^?1 s^?1. This enables the development of complementary inverters with a voltage gain of >16 and a large worst-case noise margin at a supply voltage of <0.6 V, while consuming less than 1 µW of power.     

Unraveling the Electrochemical Electrode Coupling in Integrated Organic Electrochemical Transistors

Organic electrochemical transistors (OECTs) have gained enormous attention due to their potential for bioelectronics and neuromorphic computing. However, their implementation into real-world applications is still impeded by a lack of understanding of the complex operation of integrated OECTs. This study, for the first time, elaborates on a peculiar behavior that integrated OECTs exhibit due to their electrolytic environment—the electrochemical electrode coupling (EEC), which has severe implications on the device and circuit performance, causing a loss of output saturation and a threshold voltage roll-off. After developing a physical model to describe this effect, it is substantiated with experimental data, and the crucial role of the gate electrode is discussed. Furthermore, the impact of the electrode/channel overlap on the saturation in the output curve is evaluated. It is then investigated how its detrimental effect on circuit performance can be minimized, and the optimization of a simple logic gate is demonstrated. This study has fundamental implications for researchers exploring materials and device architectures for OECTs and for engineers designing integrated OECT-based circuits.     

Versatile use of ytterbium(III) triflate and acid washed molecular sieves in the activation of glycosyl trifluoroacetimidate donors. Assemblage of a biologically relevant tetrasaccharide sequence of Globo H

The nonreducing tetrasaccharide terminus of Globo H has been assembled in good yield and excellent stereocontrol exclusively by using mild and moisture stable agents such as Yb(OTf)(3) and acid washed molecular sieves for the activation of glycosyltrifluoroacetimidate donors in the glycosylation steps.     

Synthesis of enantiomerically pure α-ethyl, α-vinyl and α-ethynyl 3,3,3-trifluoro alaninates

We describe the first synthesis of enantiomerically pure (S)-α-vinyl and (S)-α-ethynyl 3,3,3-trifluoro alanine (TF-Ala) ethyl esters, starting from the in situ generated sulfinimine (S)-2 as key fluorinated building-block. (S)-α-Ethyl TF-Ala-OEt has been prepared as well by this route.     

Native chemical ligation through in situ O to S acyl shift

[reaction: see text] A novel strategy to generate thioester peptides compatible with Fmoc chemistry is presented. Peptide-C(alpha)oxy-(2-mercapto-1-carboxyamide)ethyl ester undergoes an O to S acyl shift during ligation and the newly formed thioester intermediate reacts with an N-terminal cysteine fragment generating a product with native amide bond at the ligation site.     

Nanoscale structural and electronic properties of ultrathin blends of two polyaromatic molecules: a Kelvin probe force microscopy investigation.

We describe a Kelvin Probe Force Microscopy (KPFM) study on the morphological and electronic properties of complex mono and bi-molecular ultrathin films self-assembled on mica. These architectures are made up from an electron-donor (D), a synthetic all-benzenoid polycyclic aromatic hydrocarbon, and an electron-acceptor (A), perylene-bis-dicarboximide. The former molecule self-assembles into fibers in single component films, while the latter molecule forms discontinuous layers. Taking advantage of the different solubility and self-organizing properties of the A and D molecules, multicomponent ultrathin films characterized by nanoscale phase segregated fibers of D embedded in a discontinuous layer of A are formed. The direct estimation of the surface potential, and consequently the local workfunction from KPFM images allow a comparison of the local electronic properties of the blend with those of the monocomponent films. A change in the average workfunction values of the A and D nanostructures in the blend occurs which is primarily caused by the intimate contact between the two components and the molecular order within the nanostructure self-assembled at the surface. Additional roles can be ascribed to the molecular packing density, to the presence of defects in the film, to the different conformation of the aliphatic peripheral chains that might cover the conjugated core and to the long-range nature of the electrostatic interactions employed to map the surface by KPFM limiting the spatial and potential resolution. The local workfunction studies of heterojunctions can be of help to tune the electronic properties of active multicomponent films, which is crucial for the fabrication of efficient organic electronic devices as solar cells.