PGRP-LB: An Inside View into the Mechanism of the Amidase Reaction
| Autor: | Anna Zaidman-Rémy, Carole Vincent-Monégat, Pierre Aller, Allen M. Orville, Abdelaziz Heddi, A. Butryn, Pedro Da Silva, Laurent Soulère, Julien Orlans, Catherine Sivignon, Isabelle Rahioui |
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| Přispěvatelé: | Biologie Fonctionnelle, Insectes et Interactions (BF2I), Institut National des Sciences Appliquées de Lyon (INSA Lyon), Université de Lyon-Institut National des Sciences Appliquées (INSA)-Université de Lyon-Institut National des Sciences Appliquées (INSA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), DIAMOND Light source, Research Complex at Harwell, Institut de Chimie et Biochimie Moléculaires et Supramoléculaires (ICBMS), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National des Sciences Appliquées de Lyon (INSA Lyon), Université de Lyon-Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut de Chimie du CNRS (INC)-École Supérieure Chimie Physique Électronique de Lyon-Centre National de la Recherche Scientifique (CNRS), ANR-17-CE20-0031,GREEN,Comprendre les mécanismes de régulation et la fonction des gènes de la réponse immunitaire de l'hôte pour perturber la symbiose et le contrôle des endosymbiotes chez les insectes nuisibles(2017) |
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
Models
Molecular 0301 basic medicine QH301-705.5 Mutant Peptidoglycan Article Protein Structure Secondary Catalysis Amidohydrolases Amidase Inorganic Chemistry 03 medical and health sciences chemistry.chemical_compound Tracheal cytotoxin Amidase activity Animals Protein Isoforms peptidoglycan recognition protein Amino Acid Sequence Virulence Factors Bordetella Biology (General) Physical and Theoretical Chemistry Site-directed mutagenesis QD1-999 Drosophila melanogaster Molecular Biology innate immunity Spectroscopy X-ray crystallography Innate immune system 030102 biochemistry & molecular biology Organic Chemistry Pattern recognition receptor General Medicine Computer Science Applications PGRP-LB Zinc Chemistry 030104 developmental biology chemistry Biochemistry Mutant Proteins Carrier Proteins Sugars [SDV.EE.IEO]Life Sciences [q-bio]/Ecology environment/Symbiosis |
| Zdroj: | International Journal of Molecular Sciences International Journal of Molecular Sciences, MDPI, 2021, 22 (9), pp.4957. ⟨10.3390/ijms22094957⟩ Volume 22 Issue 9 International Journal of Molecular Sciences, Vol 22, Iss 4957, p 4957 (2021) |
| ISSN: | 1661-6596 1422-0067 |
| DOI: | 10.3390/ijms22094957⟩ |
| Popis: | Peptidoglycan recognition proteins (PGRPs) are ubiquitous among animals and play pivotal functions in insect immunity. Non-catalytic PGRPs are involved in the activation of immune pathways by binding to the peptidoglycan (PGN), whereas amidase PGRPs are capable of cleaving the PGN into non-immunogenic compounds. Drosophila PGRP-LB belongs to the amidase PGRPs and downregulates the immune deficiency (IMD) pathway by cleaving meso-2,6-diaminopimelic (meso-DAP or DAP)-type PGN. While the recognition process is well analyzed for the non-catalytic PGRPs, little is known about the enzymatic mechanism for the amidase PGRPs, despite their essential function in immune homeostasis. Here, we analyzed the specific activity of different isoforms of Drosophila PGRP-LB towards various PGN substrates to understand their specificity and role in Drosophila immunity. We show that these isoforms have similar activity towards the different compounds. To analyze the mechanism of the amidase activity, we performed site directed mutagenesis and solved the X-ray structures of wild-type Drosophila PGRP-LB and its mutants, with one of these structures presenting a protein complexed with the tracheal cytotoxin (TCT), a muropeptide derived from the PGN. Only the Y78F mutation abolished the PGN cleavage while other mutations reduced the activity solely. Together, our findings suggest the dynamic role of the residue Y78 in the amidase mechanism by nucleophilic attack through a water molecule to the carbonyl group of the amide function destabilized by Zn2+. |
| Databáze: | OpenAIRE |
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