A5033903587- Mass Spectrometry Techniques and Applications
- Enzyme Structure and Function
- Virus-based gene therapy research
- Immune Cell Function and Interaction
- Photoreceptor and optogenetics research
- Monoclonal and Polyclonal Antibodies Research
- Plant and Biological Electrophysiology Studies
- Infant Nutrition and Health
- Enzyme Catalysis and Immobilization
- Glycosylation and Glycoproteins Research
- RNA and protein synthesis mechanisms
- Biochemical and Structural Characterization
- Histone Deacetylase Inhibitors Research
- Plant biochemistry and biosynthesis
- Epigenetics and DNA Methylation
- Advanced Proteomics Techniques and Applications
- X-ray Spectroscopy and Fluorescence Analysis
- Advanced X-ray Imaging Techniques
- Genetic and Kidney Cyst Diseases
- Diet and metabolism studies
- Immunotherapy and Immune Responses
- T-cell and B-cell Immunology
- GDF15 and Related Biomarkers
- Cervical Cancer and HPV Research
- Proteoglycans and glycosaminoglycans research
Czech Academy of Sciences, Institute of Microbiology
2013-2024
Czech Academy of Sciences
2014-2024
Leibniz Institute of Virology (LIV)
2019-2024
Centre for Structural Systems Biology
2024
Institute of Virology of the Slovak Academy of Sciences
2024
Deutsches Elektronen-Synchrotron DESY
2024
European X-Ray Free-Electron Laser
2019-2023
Leibniz Association
2020-2021
Charles University
2010-2020
Département de Chimie Moléculaire
2010
Hydrogen/deuterium exchange coupled to mass spectrometry (HXMS) utilizes enzymatic digestion of proteins localize the information about altered patterns in protein structure. The ability protease produce small peptides and overlapping fragments provide sufficient coverage sequence is essential for localizing regions interest. Recently, it was shown that there an interesting group proteolytic enzymes from carnivorous pitcher plants genus Nepenthes. In this report, we describe successful...
The pitcher secretions of the Nepenthes genus carnivorous plants contain a proteolytic activity that is very useful for hydrogen/deuterium exchange mass spectrometry (HX-MS). Our efforts to reconstitute fluid using recombinant nepenthesin I (one two known aspartic proteases in fluid) revealed partial cleavage profile and reduced enzymatic stability certain HX-MS applications. We produced characterized II determine if it complemented Nepenthesin shares many properties with I, such as fast...
Eukaryotic protein homeostasis (proteostasis) is largely dependent on the action of highly conserved Hsp70 molecular chaperones. Recent evidence indicates that, apart from allostery, proteins have retained and adapted ability to assemble as functionally relevant ATP-bound dimers throughout evolution. Here, we compared ATP-dependent dimerization DnaK, human stress-inducible Hsp70, Hsc70 BiP proteins, showing that their propensities differ, with being predominantly dimeric in presence ATP....
Gas-phase activation and dissociation studies of biomolecules, proteins their non-covalent complexes using X-rays hold great promise for revealing new insights into the structure function biological samples. This is due to unique properties X-ray molecular interactions, such as site-specific rapid ionization. In this perspective, we report discuss first proof-of-principle X-ray-induced native samples ranging from small 17 kDa monomeric up large 808 protein assemblies conducted at a...
Primary hyperoxaluria type I (PH1) is caused by deficient alanine:glyoxylate aminotransferase (AGT) activity. PH1‐causing mutations in AGT lead to protein mistargeting and aggregation. Here, we use hydrogen‐deuterium exchange (HDX) characterize the wild‐type (WT), LM (a polymorphism frequent PH1 patients) G170R (the most common mutation PH1) variants of AGT. We provide first experimental analysis structural dynamics, showing that stability heterogeneous native state providing a blueprint for...
Insight into the structure–function relationship of membrane proteins is important to understand basic cell function and inform drug development, as these are common targets for drugs. Hydrogen/deuterium exchange mass spectrometry (HDX-MS) an established technique study protein conformational dynamics has shown compatibility with proteins. However, digestion analysis peptides from can be challenging, severely limiting HDX-MS experiment. Here we compare four integral proteins—Cl–/H+...
The natural function of cellobiose dehydrogenase (CDH) to donate electrons from its catalytic flavodehydrogenase (DH) domain via cytochrome (CYT) lytic polysaccharide monooxygenase (LPMO) is an example a highly efficient extracellular electron transfer chain. To investigate the CYT movement in two occurring steps, CDHs ascomycete Neurospora crassa (NcCDHIIA and NcCDHIIB) five chimeric CDH enzymes created by swapping were studied combination with fungus' own LPMOs (NcLPMO9C NcLPMO9F). Kinetic...
Article22 December 2021Open Access Source DataTransparent process Caldendrin and myosin V regulate synaptic spine apparatus localization via ER stabilization in dendritic spines Anja Konietzny RG Optobiology, Institute of Biology, Humboldt Universität zu Berlin, Germany Guest Group Neuronal Protein Transport, Center for Molecular Neurobiology, ZMNH, University Medical Hamburg-Eppendorf, Hamburg, Search more papers by this author Jasper Grendel Alan Kadek orcid.org/0000-0002-7953-5870 Leibniz...
ABSTRACT Adenovirus (AdV) infection of the respiratory epithelium is common but poorly understood. Human AdV species C types, such as HAdV-C5, utilize Coxsackie-adenovirus receptor (CAR) for attachment and subsequently integrins entry. CAR are however located deep within tight junctions in mucosa where they would not be easily accessible. Recently, a model CAR-independent entry was proposed. In this model, human lactoferrin (hLF), an innate immune protein, aids viral uptake into epithelial...
Cellobiose dehydrogenase (CDH) from wood degrading fungi represents a subclass of oxidoreductases with unique properties. Consisting two domains exhibiting interdomain electron transfer, this is the only known flavocytochrome involved in degradation. High resolution structures separated were solved, but overall architecture intact protein and exact interface unknown. Recently, it was shown that divalent cations modulate activity CDH its pH optimum possible mechanism involving bridging...
Phosducin (Pdc), a highly conserved phosphoprotein involved in the regulation of retinal phototransduction cascade, transcriptional control, and modulation blood pressure, is controlled phosphorylation-dependent manner, including binding to 14-3-3 protein. However, molecular mechanism this largely unknown. Here, solution structure Pdc its interaction with protein were investigated using small angle x-ray scattering, time-resolved fluorescence spectroscopy, hydrogen-deuterium exchange coupled...
Pentapeptide diacidic sequence LELTE, derived from the mycobacterial heat shock protein hsp65, has been recently identified as a "danger" signal of immune system effective via specific binding to universal leukocyte triggering receptor CD69. This is not active per se, only after its presentation within multivalent environment parent protein, or artificial dimerization using standard bifunctional reagents. Here we describe an entirely new way presenting this peptide based on attachment...
Abstract SARS-CoV-2 infection is associated with increased morbidities in men compared to women. Androgens are believed play an important role pathogenesis due the postulated androgen-dependency of ACE2 and TMPRSS2. However, it yet unclear whether sex bias mediated by itself or other confounding factors. Here, using golden hamster model, we show that attacks reproductive organs, causes massive dysregulation hormones induces elevated transcription androgen-to-estrogen converting enzyme...
Abstract High-risk human papillomaviruses (HPVs) cause various cancers. While type-specific prophylactic vaccines are available, additional anti-viral strategies highly desirable. Initial HPV cell entry involves receptor-switching induced by structural capsid modifications. These modifications initiated interactions with cellular heparan sulphates (HS), however, their molecular nature and functional consequences remain elusive. Combining virological assays hydrogen/deuterium exchange mass...
MS SPIDOC is a novel sample delivery system designed for single (isolated) particle imaging at X-ray Free-Electron Lasers that adaptable towards most large-scale facility beamlines. Biological samples can range from small proteins to MDa particles. Following nano-electrospray ionization, ionic be m/z-filtered and structurally separated before being oriented the interaction zone. Here, we present simulation package developed alongside this prototype. The first part describes how front-to-end...
Abstract The combination of a linear quadrupole ion-filter and Paul trap operated with rectangular guiding field for the filtering accumulation ions within M ass S pectrometry ingle P article I maging D ipole O riented protein C omplexes (MS SPIDOC) prototype [T. Kierspel et al., Anal. Bioanal. Chem. , published online] is characterized. Using cationic caesium-iodide clusters, ion-separation performance, ion accumulation, cooling, ejection via in-trap pin electrodes evaluated. Furthermore,...
Nepenthesins are aspartic proteases secreted by carnivorous pitcher plants of the genus Nepenthes. They significantly differ in sequence from other plant proteases. This difference, which provides more cysteine residues structure nepenthesins, may contribute to their unique stability profile. Recombinantly produced nepenthesin 1 (rNep1) N. gracilis complex with pepstatin A was crystallized under two different crystallization conditions using a newly formulated low-pH screen. The diffraction...