Title |
Membrane Topology and Heme Binding of the Histidine Kinases HrrS and ChrS in Corynebacterium glutamicum
|
---|---|
Published in |
Frontiers in Microbiology, February 2018
|
DOI | 10.3389/fmicb.2018.00183 |
Pubmed ID | |
Authors |
Marc Keppel, Eva Davoudi, Cornelia Gätgens, Julia Frunzke |
Abstract |
The HrrSA and the ChrSA two-component systems play a central role in the coordination of heme homeostasis in the Gram-positive soil bacteriumCorynebacterium glutamicumand the prominent pathogenCorynebacterium diphtheriae, both members of theCorynebacteriaceae. In this study, we have performed a comparative analysis of the membrane topology and heme-binding characteristics of the histidine kinases HrrS and ChrS ofC. glutamicum. While the cytoplasmic catalytic domains are highly conserved between HrrS and ChrS, the N-terminal sensing parts share only minor sequence similarity. PhoA and LacZ fusions of the N-terminal sensor domains of HrrS and ChrS revealed that both proteins are embedded into the cytoplasmic membrane via six α-helices. Although the overall membrane topology appeared to be conserved, target gene profiling indicated a higher sensitivity of the ChrS system to low heme levels (< 1 μM).In vitro, solubilized and purified full-length proteins bound heme in a 1:1 stoichiometry per monomer. Alanine-scanning of conserved amino acid residues in the N-terminal sensor domain revealed three aromatic residues (Y112, F115, and F118), which apparently contribute to heme binding of HrrS. Exchange of either one or all three residues resulted in an almost abolished heme binding of HrrSin vitro. In contrast, ChrS mutants only displayed a red shift of the soret band from 406 to 418 nm suggesting an altered set of ligands in the triple mutant. In line with target gene profiling, thesein vitrostudies suggest distinct differences in the heme-protein interface of HrrS and ChrS. Since the membrane topology mapping displayed no extensive loop regions and alanine-scanning revealed potential heme-binding residues in α-helix number four, we propose an intramembrane sensing mechanism for both proteins. Overall, we present a first comparative analysis of the ChrS and HrrS kinases functioning as transient heme sensors in theCorynebacteriaceae. |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 15 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Student > Master | 5 | 33% |
Researcher | 3 | 20% |
Student > Doctoral Student | 2 | 13% |
Student > Ph. D. Student | 2 | 13% |
Professor | 1 | 7% |
Other | 1 | 7% |
Unknown | 1 | 7% |
Readers by discipline | Count | As % |
---|---|---|
Agricultural and Biological Sciences | 5 | 33% |
Biochemistry, Genetics and Molecular Biology | 5 | 33% |
Veterinary Science and Veterinary Medicine | 1 | 7% |
Computer Science | 1 | 7% |
Engineering | 1 | 7% |
Other | 0 | 0% |
Unknown | 2 | 13% |