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Structural information about protein–RNA complexes supports the understanding of crucial recognition processes in the cell, and it can allow the development of high affinity ligands to interfere with these processes. In this respect, the identification of amino acid hotspots is particularly important. In contrast to protein–protein interactions, in silico approaches for protein–RNA interactions lag behind in their development. Herein, we report an analysis of available protein–RNA structures. We assembled a data set of 322 crystal and NMR structures and analyzed them regarding interface properties. In addition, we describe a computational alanine-scanning approach which provides interaction scores for interface amino acids, allowing the identification of potential hotspots in protein–RNA interfaces. We have made the computational approach available as an online tool, which allows interaction scores to be calculated for any structure of a protein–RNA complex by uploading atomic coordinates to the PRI HotScore web server (https://pri-hotscore.labs.vu.nl).

Advances in protein engineering have made the industrial use of enzymes feasible, though, a more wide-spread application is hampered by the limited stability of enzymes under often required harsh conditions. We developed a protein stabilization technology that involves the in situ cyclization of proteins (INCYPRO) and facilitates the design and semi-synthesis of multicyclic enzymes. These synthetically enhanced durable enzymes add to the current efforts towards a sustainable and green economy.

Roll with it: The quantitative analysis of specific miRNAs from biological samples is very likely to revolutionize diagnostics of human disease. A novel method for miRNA analysis employing rolling-circle amplification (RCA) can homogeneously detect miRNA, even at concentrations as low as 10 fM. The use of T4 RNA ligase 2 (T4 RnL2) at elevated temperatures enables very good discrimination of miRNAs differing by a single nucleotide.

Without labeling RNA: A new rapid assay for micro-RNA maturation was developed. The assay depends on the cleavage of unmodified pre-miRNAs and the subsequent amplification of the mature miRNA by rolling circle amplification (see accompanying scheme).

Proteins possess unique molecular recognition capabilities and enzymatic activities, features that are usually tied to a particular tertiary structure. To make use of proteins for biotechnological and biomedical purposes, it is often required to enforce their tertiary structure in order to ensure sufficient stability under the conditions inherent to the application of interest. The introduction of intramolecular crosslinks has proven efficient in stabilizing native protein folds. Herein, we give an overview of methods that allow the macrocyclization of expressed proteins, discussing involved reaction mechanisms and structural implications., Protein function is often tightly linked to a particular tertiary structure. For many protein applications it is required to stabilize the tertiary structure and prevent unfolding. Here, we summarize strategies that have been used to cyclize expressed proteins aiming for the stabilization of their native fold.