Identification and structural characterization of LytU, a unique peptidoglycan endopeptidase from the lysostaphin family

6533b86cfe1ef96bd12c8279

RESEARCH PRODUCT

Identification and structural characterization of LytU, a unique peptidoglycan endopeptidase from the lysostaphin family

Olli Aitio Helena Tossavainen Perttu Permi Perttu Permi Jarmo T. Juuti Vytas Raulinaitis Keiichi Hiramatsu Vesa P. Kontinen Vesa P. Kontinen

subject

0301 basic medicine entsyymit antimicrobial compounds PROTEIN chemistry.chemical_compound Catalytic Domain CELL-WALL BINDING Multidisciplinary ACTIVE-SITE Q RESISTANT STAPHYLOCOCCUS-AUREUS R Hydrogen-Ion Concentration Anti-Bacterial Agents Zinc Biochemistry Medicine HISTIDINES Protein Binding Staphylococcus aureus Science enzymes Biology Cleavage (embryo)metalloproteinases Article Cofactor BACILLUS-SUBTILIS Cell wall Structure-Activity Relationship 03 medical and health sciences Endopeptidases Protein Interaction Domains and Motifs Amino Acid Sequence staphylococci antimikrobiset yhdisteet Binding Sites Lysostaphin Cell Membrane Active site Isothermal titration calorimetry Periplasmic space VANCOMYCIN stafylokokit metalloproteinaasit MODEL 030104 developmental biology RESOLUTION chemistry Mutation Proteolysis Lysostaphin biology.protein 1182 Biochemistry cell and molecular biology Peptidoglycan

description

AbstractWe introduce LytU, a short member of the lysostaphin family of zinc-dependent pentaglycine endopeptidases. It is a potential antimicrobial agent for S. aureus infections and its gene transcription is highly upregulated upon antibiotic treatments along with other genes involved in cell wall synthesis. We found this enzyme to be responsible for the opening of the cell wall peptidoglycan layer during cell divisions in S. aureus. LytU is anchored in the plasma membrane with the active part residing in the periplasmic space. It has a unique Ile/Lys insertion at position 151 that resides in the catalytic site-neighbouring loop and is vital for the enzymatic activity but not affecting the overall structure common to the lysostaphin family. Purified LytU lyses S. aureus cells and cleaves pentaglycine, a reaction conveniently monitored by NMR spectroscopy. Substituting the cofactor zinc ion with a copper or cobalt ion remarkably increases the rate of pentaglycine cleavage. NMR and isothermal titration calorimetry further reveal that, uniquely for its family, LytU is able to bind a second zinc ion which is coordinated by catalytic histidines and is therefore inhibitory. The pH-dependence and high affinity of binding carry further physiological implications.

year	journal	country	edition	language
2017-07-01	Scientific Reports

10.1038/s41598-017-06135-w https://doaj.org/article/c8e8573d85a24ba684ea4c55fbb932a8