The ubiquitin-proteasome system (UPS) has been successfully targeted by both academia and the pharmaceutical industry for oncological and immunological applications. Typical proteasome inhibitors are based on a peptidic backbone endowed with an electrophilic C-terminus by which they react with the active proteolytic sites. Although the peptide moiety has attracted much attention in terms of subunit selectivity, the target specificity and biological stability of the compounds are largely determined by the reactive warheads. In this study, we have carried out a systematic investigation of described electrophiles by a combination of in vitro, in vivo, and structural methods in order to disclose the implications of altered functionality and chemical reactivity. Thereby, we were able to introduce and characterize the class of α-ketoamides as the most potent reversible inhibitors with possible applications for the therapy of solid tumors as well as autoimmune disorders.

The problem how to approximately determine the absolute value of the Fisher information measure for a general parametric probabilistic system is considered. Having available the first and second moment of the system output in a parametric form, it is shown that the information measure can be bounded from below through a replacement of the original system by a Gaussian system with equivalent moments. The presented technique is applied to a system of practical importance and the potential quality of the bound is demonstrated.Comment: 4 pages, submitted to IEEE Signal Processing Letter

Natural products represent valuable lead structures for drug discovery. However, for most bioactive compounds no cellular target is yet identified and many substances predicted from genome analysis are inaccessible due to their life stage-dependent biosynthesis, which is not reflected in common isolation procedures. In response to these issues, an NMR-based and target-directed protease assay for inhibitor detection of the proteasome was developed. The methodology is suitable for one-shot identification of inhibitors in conglomerates and crude culture broths. The technique was applied for analysis of the different life stages of the bacterium Photorhabdus luminescens, which resulted in the isolation and characterization of cepafungin I (CepI), the strongest proteasome inhibitor described to date. Its biosynthesis is strictly regulated and solely induced by the specific environmental conditions determined by our methodology. The transferability of the developed technique to other drug targets may disclose an abundance of novel compounds applicable for drug development.NMR assay | UPS | natural compound | screening | virulence

Small heat shock proteins (sHsps) are ubiquitous molecular chaperones that prevent the aggregation of unfolding proteins during proteotoxic stress. In Caenorhabditis elegans, Sip1 is the only sHsp exclusively expressed in oocytes and embryos. Here, we demonstrate that Sip1 is essential for heat shock survival of reproducing adults and embryos. X-ray crystallography and electron microscopy revealed that Sip1 exists in a range of well-defined globular assemblies consisting of two half-spheres, each made of dimeric "spokes." Strikingly, the oligomeric distribution of Sip1 as well as its chaperone activity depend on pH, with a trend toward smaller species and higher activity at acidic conditions such as present in nematode eggs. The analysis of the interactome shows that Sip1 has a specific substrate spectrum including proteins that are essential for embryo development.

A number of 2-(2-hydroxyphenyl)-4,6-diaryl-1,3,5-triazines (HPTs) and TIN P (2-(2-hydroxy-5-methylphenyl)benzotriazole) show phosphorescence in polar solvents at 77 K which increases in intensity with UV-irradiation time until an equilibrium value is reached (phosphorescence evolution). TIN P phosphoresces even at the very beginning of irradiation, in contrast to the HPTs, such as M-OH-P (2-(2-hydroxy-4-methoxyphenyl)-4,6-diphenyl-1,3,5-triazine), which exhibit no such initial phosphorescence provided that they were not recently exposed to UV radiation. The corresponding methoxy derivatives (MPTs) of some HPTs, where the H atom of the intramolecular hydrogen bond (IMHB) is replaced by a methyl group, produce intense phosphorescence independent of irradiation time. Considerable relaxation is found for HPTs after dark periods ≤ 1 h at 77 K resulting in a significantly lower initial phosphorescence intensity upon renewed irradiation. TIN P, in contrast, shows much slower relaxation which becomes significant only at elevated temperatures. Phosphorescence evolution is due to open conformers of the molecules, i.e., with intermolecular rather than intramolecular hydrogen bonds, which are formed in polar solvents under the influence of UV radiation. Relaxation, i.e., re-formation of the IMHB of open-form molecules, is faster for the investigated HPTs than for TIN P.

Caseinolytic protease P (ClpP) is an important regulator of Staphylococcus aureus pathogenesis. A high-throughput screening for inhibitors of ClpP peptidase activity led to the identification of the first non-covalent binder for this enzyme class. Co-crystallization of the small molecule with S. aureus ClpP revealed a novel binding mode: Because of the rotation of the conserved residue proline 125, ClpP is locked in a defined conformational state, which results in distortion of the catalytic triad and inhibition of the peptidase activity. Based on these structural insights, the molecule was optimized by rational design and virtual screening, resulting in derivatives exceeding the potency of previous ClpP inhibitors. Strikingly, the conformational lock is overturned by binding of ClpX, an associated chaperone that enables proteolysis by substrate unfolding in the ClpXP complex. Thus, regulation of inhibitor binding by associated chaperones is an unexpected mechanism important for ClpP drug development.

No abstract

In this work, the problem of signal parameter estimation from measurements acquired by a low-complexity analog-to-digital converter (ADC) with 1-bit output resolution and an unknown quantization threshold is considered. Singlecomparator ADCs are energy-efficient and can be operated at ultra-high sampling rates. For analysis of such systems, a fixed and known quantization threshold is usually assumed. In the symmetric case, i.e., zero hard-limiting offset, it is known that in the low signal-to-noise ratio (SNR) regime the signal processing performance degrades moderately by 2/π (−1.96 dB) when comparing to an ideal ∞-bit converter. Due to hardware imperfections, low-complexity 1-bit ADCs will in practice exhibit an unknown threshold different from zero. Therefore, we study the accuracy which can be obtained with receive data processed by a hard-limiter with unknown quantization level by using asymptotically optimal channel estimation algorithms. To characterize the estimation performance of these nonlinear algorithms, we employ analytic error expressions for different setups while modeling the offset as a nuisance parameter. In the low SNR regime, we establish the necessary condition for a vanishing loss due to missing offset knowledge at the receiver. As an application, we consider the estimation of single-input single-output wireless channels with inter-symbol interference and validate our analysis by comparing the analytic and experimental performance of the studied estimation algorithms. Finally, we comment on the extension to multiple-input multiple-output channel models.