Determination of kinetics and crystal structure of a novel Type 2 Isopentenyl Diphosphate: Dimethylallyl Diphosphate Isomerase from Streptococcus pneumoniae
| Autor: | Syam Sundar Neti, Johan Wouters, Heidi L. Schubert, C. Dale Poulter, Rita M. Cornish, Jérôme de Ruyck, Matthew Walter Janczak, André Matagne, Steven Cary Rothman |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2014 |
| Předmět: |
Models
Molecular Stereochemistry Protein Conformation Flavoprotein bisubstrates Flavin group Isomerase medicine.disease_cause Crystallography X-Ray Biochemistry Enterococcus faecalis Cofactor Article Pneumococcal Infections Hemiterpenes steady-state kinetics Catalytic Domain medicine Humans Cloning Molecular Molecular Biology Escherichia coli chemistry.chemical_classification flavoproteins isoprenoids biology Chemistry Organic Chemistry Active site biology.organism_classification Carbon-Carbon Double Bond Isomerases Enzyme Streptococcus pneumoniae Drug Design biology.protein Molecular Medicine X-ray structures |
| Popis: | Isopentenyl diphosphate isomerase (IDI) is a key enzyme in the isoprenoid biosynthetic pathway and is required for all organisms that synthesize isoprenoid metabolites from mevalonate. Type 1 IDI (IDI-1) is a metalloprotein that is found in eukaryotes, whereas the type 2 isoform (IDI-2) is a flavoenzyme found in bacteria that is completely absent from human. IDI-2 from the pathogenic bacterium Streptococcus pneumoniae was recombinantly expressed in Escherichia coli. Steady-state kinetic studies of the enzyme indicated that FMNH2 (KM=0.3 μM) bound before isopentenyl diphosphate (KM=40 μM) in an ordered binding mechanism. An X-ray crystal structure at 1.4 Å resolution was obtained for the holoenzyme in the closed conformation with a reduced flavin cofactor and two sulfate ions in the active site. These results helped to further approach the enzymatic mechanism of IDI-2 and, thus, open new possibilities for the rational design of antibacterial compounds against sequence-similar and structure-related pathogens such as Enterococcus faecalis or Staphylococcus aureus. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|