Keyword search (3,448 papers available)


A shared binding site for NAD+ and coenzyme A in an acetaldehyde dehydrogenase involved in bacterial degradation of aromatic compounds.

Author(s): Lei Y, Pawelek PD, Powlowski J

Biochemistry. 2008 Jul 01;47(26):6870-82 Authors: Lei Y, Pawelek PD, Powlowski J

Article GUID: 18537268
Subunit orientation in the Escherichia coli enterobactin biosynthetic EntA-EntE complex revealed by a two-hybrid approach.

Author(s): Pakarian P, Pawelek PD

Biochimie. 2016 Aug;127:1-9 Authors: Pakarian P, Pawelek PD

Article GUID: 27086082
Intracellular co-localization of the Escherichia coli enterobactin biosynthetic enzymes EntA, EntB, and EntE.

Author(s): Pakarian P, Pawelek PD

Biochem Biophys Res Commun. 2016 09 09;478(1):25-32 Authors: Pakarian P, Pawelek PD

Article GUID: 27470582
A novel set of vectors for Fur-controlled protein expression under iron deprivation in Escherichia coli.

Author(s): Pakarian P, Pawelek PD

BMC Biotechnol. 2016 09 13;16(1):68 Authors: Pakarian P, Pawelek PD

Article GUID: 27619907
Dual Activity of Rose Bengal Functionalized to Albumin-Coated Lanthanide-Doped Upconverting Nanoparticles: Targeting and Photodynamic Therapy.

Author(s): Sabri T, Pawelek PD, Capobianco JA

ACS Appl Mater Interfaces. 2018 Aug 15;10(32):26947-26953 Authors: Sabri T, Pawelek PD, Capobianco JA

Article GUID: 30028124
Title:Intracellular co-localization of the Escherichia coli enterobactin biosynthetic enzymes EntA, EntB, and EntE.
Authors:Pakarian P,  Pawelek PD
Link:https://www.ncbi.nlm.nih.gov/pubmed/27470582?dopt=Abstract
Category:Biochem Biophys Res Commun
PMID:27470582
Dept Affiliation: CHEMBIOCHEM
1 Department of Chemistry and Biochemistry, Concordia University, 7141 Sherbrooke St., W., Montreal, H4B 1R6, Quebec, Canada.
2 Department of Chemistry and Biochemistry, Concordia University, 7141 Sherbrooke St., W., Montreal, H4B 1R6, Quebec, Canada; Groupe de Recherche Axé sur la Structure des Protéines (GRASP), Canada. Electronic address: peter.pawelek@concordia.ca.

Description:

Intracellular co-localization of the Escherichia coli enterobactin biosynthetic enzymes EntA, EntB, and EntE.

Biochem Biophys Res Commun. 2016 09 09;478(1):25-32

Authors: Pakarian P, Pawelek PD

Abstract

Bacteria utilize small-molecule iron chelators called siderophores to support growth in low-iron environments. The Escherichia coli catecholate siderophore enterobactin is synthesized in the cytoplasm upon iron starvation. Seven enzymes are required for enterobactin biosynthesis: EntA-F, H. Given that EntB-EntE and EntA-EntE interactions have been reported, we investigated a possible EntA-EntB-EntE interaction in E. coli cells. We subcloned the E. coli entA and entB genes into bacterial adenylate cylase two-hybrid (BACTH) vectors allowing for co-expression of EntA and EntB with N-terminal fusions to the adenylate cyclase fragments T18 or T25. BACTH constructs were functionally validated using the CAS assay and growth studies. Co-transformants expressing T18/T25-EntA and T25/T18-EntB exhibited positive two-hybrid signals indicative of an intracellular EntA-EntB interaction. To gain further insights into the interaction interface, we performed computational docking in which an experimentally validated EntA-EntE model was docked to the EntB crystal structure. The resulting model of the EntA-EntB-EntE ternary complex predicted that the IC domain of EntB forms direct contacts with both EntA and EntE. BACTH constructs that expressed the isolated EntB IC domain fused to T18/T25 were prepared in order to investigate interactions with T25/T18-EntA and T25/T18-EntE. CAS assays and growth studies demonstrated that T25-IC co-expressed with the EntB ArCP domain could complement the E. coli entB(-) phenotype. In agreement with the ternary complex model, BACTH assays demonstrated that the EntB IC domain interacts with both EntA and EntE.

PMID: 27470582 [PubMed - indexed for MEDLINE]