InfraLife call to promote infrastructure use and collaborative projects.

Crystallographic fragment screening combined with in silico design to identify novel antivirals

Flavio Ballante, Karolinska Institutet

This project will establish a collaboration between the crystallographic fragment screening facility at MAX IV (FragMAX) and the Chemical Biology Consortium Sweden (CBCS, SciLifeLab), with the aim of developing integrated workflows for structure-based drug design. In a pilot project, the facilities will partner with Jens Carlsson’s group (JC, Uppsala University) to develop inhibitors targeting enzymes involved in flavivirus replication. Direct-acting antivirals are urgently needed to combat future pandemics caused by Zika or Dengue viruses. By combining crystallographic fragment screens at FragMAX with computational chemistry, biochemical assays, and compound management at CBCS, we will rapidly identify novel inhibitors. Our initiative will create a framework for facility collaboration and leverage the Carlsson group’s experience in fragment-based drug design. The project will provide starting points for drug discovery and pave the way for joint projects between MAX IV and SciLifeLab.

Spatial biology across scales and matters – combining omics with element distribution and structure

Charlotte Stadler, KTH Royal Institute of Technology

Research groups and staff scientists at Max IV and the Infrastructure Unit Spatial Proteomics at SciLIfeLab, has initiated collaboration to integrate two technologies available at Max IV -nano X-Ray Fluorescence (nano-XRF) and optical photothermal infrared spectroscopy (OPTIR), used to map chemical elements and structural changes respectively, with multiplexed immunofluorescence solutions in the SP unit at SciLIfeLab. The combination of these modalities would go beyond what is possible with technologies in the respective infrastructure, providing additional layers of information in the intact spatial context of tissue sections and cells. The possibilities for future applications are broad and a few concrete projects following successful method integration include novel insights into chemotherapy resistance in cancer, respiratory pathology and Alzheimer’s disease.

Structural basis for the role of angiotensin-converting enzyme 2 (ACE2) as a novel regulator of transmembrane serine protease 2 (TMPRSS2) catalyzed proteolytic activation of outer structural proteins of respiratory viruses

Wolfgang Knecht, Lund University

Proteolytic cleavage of viral envelope proteins by host cell proteases is essential for the infectivity of many viruses and relevant proteases provide promising drug targets. TMPRSS2 is a major activating protease of several respiratory viruses, including influenza A virus and SARS-CoV-2. The interaction of ACE2 with TMPRSS2 has been shown previously. Prof. Eva Friebertshäuser’s group has now shown that ACE2 binds to TMPRSS2 without cleaving it but increases its enzymatic activity by an unknown mechanism. We aim to determine the structural basis of this activation. Understanding this mechanism opens a new modality to target host proteases to combat respiratory viral infections by aiding structure-based drug discovery. We also will establish hereby a collaboration between Swedish national and university research infrastructures (PPS & LP3), SciLifeLab ISB units, BioMAX beamline including FragMAX, and Prof. Friebertshäuser’s group, a world-leading group in respiratory virus research.

How the HtrA2:XIAP interplay initiates the apoptotic cascade

Björn Burmann, Gothenburg University

The human mitochondrial serine protease HtrA2 plays a key role in the cellular protein quality control but also participates in the apoptotic cascade by acting as a pro-apoptotic agent which directly binds and cleaves inhibitor of apoptosis (IAP) proteins, a process which has previously been shown to upregulate the proteolytic activity of HtrA2. In our ongoing work, we used advanced solution NMR spectroscopy methods combined with biophysical characterization to characterize how HtrA2 is allosterically activated upon binding of its natural substrate, X-linked inhibitor of apoptosis protein (XIAP). Despite shedding light in the functional consequences of the HtrA2:XIAP interaction the exact nature of the complex remains elusive due to the lack of structural information partially due to the large inherent flexibility of the XIAP protein. Therefore, we plan to obtain a high-resolution structural model using an integrated structural biology approach.