Antibiotic and antiviral pressure has promoted the appearance of multi-resistant pathogens, generating an urgent need for novel antimicrobials. The structural and functional characterization of pneumococcal PG hydrolases has allowed us to design new enzymes that efficiently and selectively kill Streptococcus pneumoniae as well as other Gram-positive and Gram–negative pathogens. In addition, screening of chemical libraries is being used to find new antimicrobials interfering with microbial infections to overcome current treatment failure.
Project partners: Drs. P. García (CIB-CSIC), Mª J. Camarasa (IQM-CSIC) and J. Sancho (BIFI-UNIZAR).
Vázquez R, Domenech M, Iglesias-Bexiga M, Menéndez M, García P. 2017. Csl2, a novel chimeric bacteriophage lysin to fight infections caused by Streptococcus suis, an emerging zoonotic pathogen. Sci Rep. 2017 Nov 28;7(1):16506. doi: 10.1038/s41598-017 -16736-0. PMID: 29184097.
Blázquez B, Fresco-Taboada A, Iglesias-Bexiga M, Menéndez M, García P. 2016. PL3 amidase, a tailor-made lysin constructed by domain shuffling with potent killing activity against pneumococci and related species. Front Microbiol. 7:1156. PMID:27516758
Díez-Martínez R, de Paz HD, García-Fernández E, Bustamante N, Euler CW, Fischetti VA, Menéndez M, García P. 2015. A novel chimeric phage lysin with high in vitro and in vivo bactericidal activity against Streptococcus pneumoniae. J Antimicrob Agents, 70: 1763-1773. PMID: 25733585
Moscoso M, Esteban-Torres M, Menéndez M, García E. 2014. In vitro bactericidal and bacteriolytic activity of ceragenin CSA-13 against planktonic cultures and biofilms of Streptococcus pneumoniae and other pathogenic streptococci. PLoS One. 9(7): e101037. PMID: 25006964
Díez-Martínez, R., De Paz, H., Bustamante, N., García. E., Menéndez M., García P. 2013 Improving the lethal effect of Cpl-7, a pneumococcal phage lysozyme with broad bactericidal activity, by inverting the net charge of its cell wall-binding module. Antimicrob. Agents. Chemother. 57, 5355-5365. PMID: 23959317
We develop new designer’s microarrays for the study of probe features and recognition by different receptors. Using this technology, we can incorporate, and subsequently evaluate, multiple samples in a single chip, from purified molecules to whole cells. Novel microarray platforms already developed comprise a variety of natural complex systems as probes, as cell lysates, extracellular vesicles, bacteria wall fragments or entire bacteria, as well as yeast cells. Applications of these platforms include the study of specific features of the probes, as e.g. glycosylation patterns, exploration of probe recognition by antibodies, enzymes and different types of potential counter-receptors, and evaluation of compounds as inhibitors of recognized interactions. The new microarrays also serve to investigate ligand recognition by receptors present in the probes, as for example bacterial adhesins, using when necessary tailored immobilization strategies.
Research partners: Drs. JA Bengoechea (Queen’s University) and J. Garmendia (IdAB-CSIC), and Profs. E. García (CIB-CSIC) and E. Buzás (Semmelweis University).
Kalograiaki I, Euba B, Proverbio D, Campanero-Rhodes MA, Aastrup T, Garmendia J, Solís D. 2016. Combined Bacteria Microarray and Quartz Crystal Microbalance Approach for Exploring Glycosignatures of Nontypeable Haemophilus influenzae and Recognition by Host Lectins. Anal Chem. 88(11):5950-5957. PMID:27176788
Campanero-Rhodes MA, Llobet E, Bengoechea JA, Solís D. Bacteria microarrays as sensitive tool for exploring pathogen surface epitopes and recognition by host receptors. 2015. RSC Advances, 5: 7173-7181. doi: 10.1039/C4RA14570D
Recognition of pathogen-associated molecular patterns (PAMPs) by receptors of the innate immune system triggers activation of different defense mechanisms. Using the microarray technology recently developed by the group, we investigate the recognition of different bacteria, serotypes and mutant strains by a variety of receptors, as e.g. antibodies, the complement subcomponent C1q or endogenous lectins of the innate immune system. In parallel, we also analyze the glycosylation patterns of the bacterial surface. The rationale for this study is that the absence of ligands for endogenous lectins may be related with a higher virulence of the bacteria. Therefore, the results obtained may facilitate the design of more effective vaccines, by incorporating those serotypes not recognized efficiently by the lectins of the innate immune system, and the development of new strategies based on the selective use of lectins as therapeutic agents.
Research partners: Drs. JA Bengoechea (Queen’s University), T. Aastrup (Attana), J. Garmendia (IdAB-CSIC), C. Ardanuy (Bellvitge Universitary Hospital) and Prof. E. García (CIB-CSIC).
Kalograiaki I, Campanero-Rhodes MA, Proverbio D, Euba B, Garmendia J, Aastrup T, Solís D. 2018. Bacterial Surface Glycans: Microarray and QCM Strategies for Glycophenotyping and Exploration of Recognition by Host Receptors. Methods Enzymol. 598:37-70. doi: 10.1016/bs.mie.2017.06.011. PMID: 29306443
Martínez I, Oliveros JC, Cuesta I, de la Barrera J, Ausina V, et al. 2017. Apoptosis, Toll-like, RIG-I-like and NOD-like receptors are pathways jointlyinduced by diverse respiratory bacterial and viral pathogens. Front. Microbiol. 8:276. PMID:28298903
Kalograiaki I, Euba B, Proverbio D, Campanero-Rhodes MA, Aastrup T, Garmendia J, Solís D. 2016. Combined Bacteria Microarray and Quartz Crystal Microbalance Approach for Exploring Glycosignatures of Nontypeable Haemophilus influenzae and Recognition by Host Lectins. Anal Chem. 88(11):5950-5957. PMID:27176788
Campanero-Rhodes MA, Llobet E, Bengoechea JA, Solís D. 2015. Bacteria microarrays as sensitive tool for exploring pathogen surface epitopes and recognition by host receptors. RSC Advances, 5: 7173-7181. doi: 10.1039/C4RA14570D
Coding of bioinformation in glycans and transfer of this information via lectins is key to numerous physiological and pathological processes. Characterization of the lectins’ structure/function relationships is essential to understand their biological functions and paves the way to their pharmaceutical exploitation. A main focus is placed on galectins, a family of galactoside-binding lectins that are involved in immune and inflammatory responses, cancer and other biomedically relevant processes. Other proteins recently studied are the human macrophage Gal-specific lectin (a key receptor for the Tn carcinoma associated antigen) and different scFvs antibody fragments interacting with glycotopes involved in the rejection of xenotransplants.
Research partners: Profs. H-J Gabius (LMU Munich), J Jiménez-Barbero (CIC bioGUNE), and FJ Cañada (CIB-CSIC) and Dr. M Kilcoyne (NUI Galway).
Solís D, Bovin NV, Davis AP, Jiménez-Barbero J, Romero A, Roy R, Smetana K Jr, Gabius HJ. 2015. A guide into glycosciences: How chemistry, biochemistry and biology cooperate to crack the sugar code. 2015. Biochim Biophys Acta, 1850:186-235. PMID: 24685397
Ruiz FM, Scholz BA, Buzamet E, Kopitz J, André S, Menéndez M, Romero A, Solís D, Gabius HJ. 2014. Natural single amino acid polymorphism (F19Y) in human galectin-8: detection of structural alterations and increased growth-regulatory activity on tumor cells. FEBS J. 281(5):1446-64. PMID: 24418318
Ruiz, FM, Fernández IS, López-Merino L, Lagartera L, Kaltner H, Menéndez M, André S, Solís D, Gabius H-J, Romero A. 2013. Fine-tuning of prototype chicken galectins: Structure of CG-2 and structure-activity correlations. Acta Cryst. D: Biol. Cryst. 69: 1665-1676. PMID: 23999290
Gabius H-J, André S, Jiménez-Barbero J, Romero A and Solís D. 2011. From lectin structure to functional glycomics: principles of the sugar code. Trends Biochem Sci 36: 298-313. PMID: 21458998
Streptococcus pneumoniae is a major cause of morbidity and mortality worldwide. Pneumococcal proteins involved in cell wall hydrolysis (CW hydrolases) are essential for cell expansion and division and contribute in different ways to pneumococcal virulence. In addition, the hydrolases encoded by pneumococcal bacteriophages have emerged as a promising alternative to antibiotics as new antimicrobials. In depth characterization of the bacterial and phage enzymes has provided the first complete structures and models of several family members, clues about substrate specificity and structural motifs recognized by their CW binding domains, and has demonstrated the relevance of modularity for activity. The design of mutants and/or chimeras with different selectivities or enhanced activities has thus been facilitated.
Project partners: Drs. P García and F Díaz (CIB-CSIC); JA Hermoso and M. Bruix (IQFR-CSIC).
Bustamante N, Iglesias-Bexiga M, Bernardo-García N, Silva-Martín N, García G, Campanero-Rhodes MA, García E, Usón I, Buey RM, García P, Hermoso JA, Bruix M, Menéndez M. 2017. Deciphering how Cpl-7 cell wall-binding repeats recognize the bacterial peptidoglycan. Sci Rep. 2017 Nov 28;7(1):16494. doi: 10.1038/s41598-017-16392-4. PMID: 29184076.
Rico-Lastres P, Díez-Martínez R, Iglesias-Bexiga M, Bustamante N, Aldridge C, Hesek D, Lee M, Mobashery S, Gray J, Vollmer W, García P, Menéndez M. 2015. Substrate recognition and catalysis by LytB, a pneumococcal peptidoglycan hydrolase involved in virulence. Sci. Rep., 5, 16198; doi: 10.1038/srep16198. PMID: 26537571
Bustamante N, Campillo NE, García E, Gallego C, Pera B, Diakun GP, Sáiz JL, García P, Díaz JF, Menéndez M. 2010. Cpl-7, a lysozyme encoded by a pneumococcal bacteriophage with a novel cell wall-binding motif. J Biol Chem 285: 33184-33196. PMID: 20720016
Hermoso JA, Lagartera L, González A, García P, Martínez-Ripoll M, García JL, Menéndez M. 2005. Crystal structure of the modular teichoic acid phosphorylcholine esterase Pce from Streptococcus pneumoniae: insights into the mechanism of bacterial pathogenesis. Nat Struct Mol Biol 12:533-538. PMID: 15895092