A5001849114- Photosynthetic Processes and Mechanisms
- Metal-Catalyzed Oxygenation Mechanisms
- Enzyme Structure and Function
- Metalloenzymes and iron-sulfur proteins
- Protein Structure and Dynamics
- Hemoglobin structure and function
- Cancer, Hypoxia, and Metabolism
- Porphyrin Metabolism and Disorders
- Mass Spectrometry Techniques and Applications
- Amino Acid Enzymes and Metabolism
- Enzyme Catalysis and Immobilization
- Cell death mechanisms and regulation
- RNA and protein synthesis mechanisms
- Mitochondrial Function and Pathology
- Analytical Chemistry and Sensors
- Photoreceptor and optogenetics research
- Glycosylation and Glycoproteins Research
- Electrochemical Analysis and Applications
- Metabolism and Genetic Disorders
- Microbial Metabolic Engineering and Bioproduction
- Porphyrin and Phthalocyanine Chemistry
- Electrochemical sensors and biosensors
- Biochemical and Molecular Research
- bioluminescence and chemiluminescence research
- Force Microscopy Techniques and Applications
Universidad de Zaragoza
2014-2025
Instituto de Investigación Sanitaria Aragón
2022
Centro Médico Sanitas Zaragoza
2017
Instituto de Biocomputación y Física de Sistemas Complejos
2012-2015
Consejo Superior de Investigaciones Científicas
2009
Biocom
2007
Centro de Investigaciones Científicas Isla de la Cartuja
2003
Universidad de Sevilla
2003
Instituto de Bioquímica Vegetal y Fotosíntesis
2003
European Molecular Biology Laboratory
2000
Protein O-fucosylation is an essential post-translational modification, involved in the folding of target proteins and role these during embryonic development adult tissue homeostasis, among other things. Two different enzymes are responsible for this O-fucosyltransferase 1 2 (POFUT1 POFUT2, respectively). Both have been characterised biologically enzymatically but nothing known at molecular or structural level. Here we describe first crystal structure a catalytically functional POFUT1...
The apoptosis-inducing factor (AIF) is a mitochondrial-flavoprotein that, after cell death induction, distributed to the nucleus mediate chromatinolysis. In mitochondria, AIF present in monomer-dimer equilibrium that reduction by NADH gets displaced toward dimer. crystal structure of human (hAIF):NAD(H)-bound dimer revealed one FAD and, unexpectedly, two NAD(H) molecules per protomer. A 1:2 hAIF:NAD(H) binding stoichiometry was additionally confirmed solution using surface plasmon resonance....
A combination of structural, thermodynamic, and transient kinetic data on wild-type mutant Anabaena vegetative cell ferredoxins has been used to investigate the nature protein−protein interactions leading electron transfer from reduced ferredoxin oxidized ferredoxin:NADP+ reductase (FNR). We have determined reduction potentials ferredoxin, heterocyst 12 site-specific mutants at seven surface residues as well one- two-electron FNR, both alone in complexes with three ferredoxins. X-ray...
The crystal structure of Anabaena PCC 7119 ferredoxin-NADP+ reductase (FNR) suggests that the carboxylate group Glu301 may be directly involved in catalytic process electron and proton transfer between isoalloxazine moiety FAD FNR substrates (NADPH, ferredoxin, flavodoxin). To assess this possibility, was removed by mutating residue to an alanine. Various spectroscopic techniques (UV-vis absorption, fluorescence, CD) indicate mutant protein folded properly significant structural...
The R dx A oxygen‐insensitive nitroreductase of the human gastric pathogen H elicobacter pylori is responsible for susceptibility this organism to redox active prodrug metronidazole [2‐(2‐methyl‐5‐nitro‐1 ‐imidazol‐1‐yl)ethanol]. Loss‐of‐function mutations in rdxA are primarily resistance therapeutic. exhibits potent NADPH oxidase activity under aerobic conditions and reductase strictly anaerobic conditions. In present study, we report crystal structure , which a homodimer exhibiting domain...
Plastidic ferredoxin-NADP+ reductase (FNR) transfers two electrons from ferredoxin or flavodoxin molecules to NADP+, generating NADPH. The forces holding the Anabaena FNR:NADP+ complex were analyzed by dynamic force spectroscopy, using WT FNR and three C-terminal Y303 variants, Y303S, Y303F, Y303W. was covalently immobilized on mica NADP+ attached AFM tips. Force-distance curves collected for different loading rates specific unbinding under Bell-Evans model obtain mechanostability parameters...
Abstract Background The prokaryotic FAD synthetase family – a group of bifunctional enzymes that catalyse riboflavin phosphorylation and FMN adenylylation within single polypeptide chain- was analysed in terms sequence structure. Results Sequences nearly 800 species were aligned. Those related with activities showed conservation several consensus regions highly conserved residues. A 3D model for the from Corynebacterium ammoniagene s ( Ca FADS) generated. This confirms N-terminal C-terminal...
Helicobacter pylori establishes life-long infections in the gastric mucosa of over 1 billion people worldwide. In many cases, without specific antimicrobial intervention, H. infected individuals will develop type B gastritis, chronic peptic ulcers and, more rarely, neoplasias. Conventional therapy has been complicated by dramatic increases resistance to macrolides, metronidazole and fluoroquinolones. Here, we report development novel therapeutics that specifically target unique flavodoxin...
We report a detailed experimental study of maghemite nanoparticles, with sizes ranging from 1.6 to 6 nm, synthesized inside biological mould apoferritin. The structural characterization the inorganic cores, using TEM and x-ray diffraction, reveals low degree crystalline order, possibly arising nucleation growth multiple domains each molecule. have also investigated molecular structure by means atomic force microscopy in liquid. find that synthesis nanoparticles apoferritin leads small, but...
Ferredoxin−NADP+ reductase (FNR) catalyzes the reduction of NADP+ to NADPH in an overall reversible reaction, showing some differences mechanisms between cyanobacterial and higher plant FNRs. During hydride transfer it is proposed that FNR C-terminal Tyr displaced by nicotinamide. Thus, this might be involved not only modulating flavin redox properties, as already shown, but also nicotinamide binding transfer. variants from cyanobacterium Anabaena which has been replaced Trp, Phe, or Ser...
The thermodynamics of the formation binary and ternary complexes between Anabaena PCC 7119 FNR its substrates, NADP+ Fd, or Fld, has been studied by ITC. Despite structural dissimilarities, main difference Fd Fld binding to relates hydrophobicity, reflected in different heat capacity number water molecules released from interface. At pH 8, is both enthalpically entropically driven, accompanied protonation at least one ionizable group. His299 identified as responsible for proton exchange...
Abstract Transient absorbance measurements following laser flash photolysis have been used to measure the rate constants for electron transfer (et) from reduced Anabaena ferredoxin (Fd) wild‐type and seven site‐specific charge‐reversal mutants of ferredoxin:NADP+ reductase (FNR). These mutations designed probe importance specific positively charged amino acid residues on surface FNR molecule near exposed edge FAD cofactor in protein‐protein interaction during et with Fd. The mutant proteins...
Immobilization of proteins in a functionally active form and proper orientation is fundamental for effective surface-based protein analysis. A new method presented the controlled oriented immobilization ordered monolayers enzymes whose interaction site had been protected using ligand. The utility this was demonstrated by analyzing interactions between enzyme ferredoxin-NADP+ reductase (FNR) its redox partner ferredoxin (Fd). quality procedure deeply evaluated through enzymatic assays atomic...
Riboflavin kinases (RFKs) catalyse the phosphorylation of riboflavin to produce FMN. In most bacteria this activity is catalysed by C-terminal module a bifunctional enzyme, FAD synthetase (FADS), which also catalyses transformation FMN into through its N-terminal adenylyltransferase (FMNAT) module. The RFK FADS homologue eukaryotic monofunctional RFKs, while FMNAT lacks homologyto enzymes involved in production. Previously, crystal structure Corynebacterium ammoniagenes ( Ca FADS) was...
Previous studies and the crystal structure of Anabaena PCC 7119 FNR suggest that side chains Arg100 Arg264 may be directly involved in proper NADP+/NADPH orientation for an efficient electron-transfer reaction. Protein engineering on from has been carried out to investigate their roles complex formation electron transfer NADP+ ferredoxin/flavodoxin. replaced with alanine, which removes positive charge, long chain, as well ability form hydrogen bonds, while a charge reversal mutation made at...
The temperature dependence of hydride transfer from the substrate to N5 FAD cofactor during reductive half-reaction Pleurotus eryngii aryl-alcohol oxidase (AAO) is assessed here. Kinetic isotope effects on both pre-steady state reduction enzyme and its steady-state kinetics, with differently deuterated substrates, suggest an environmentally-coupled quantum-mechanical tunnelling process. Moreover, those kinetic data, along crystallographic structure in complex a analogue, indicate that AAO...
Bifunctional FAD synthetases (FADSs) fold in two independent modules; The C-terminal riboflavin kinase (RFK) catalyzes the RFK activity, while N-terminal FMN-adenylyltransferase (FMNAT) exhibits FMNAT activity. search for macromolecular interfaces Corynebacterium ammoniagenes FADS (CaFADS) crystal structure predicts a dimer of trimers organization. Within each trimer, head-to-tail arrangement causes and catalytic sites neighboring protomers to approach, agreement with active site residues...