Novel protein interactions with an actin homolog (MreB) of Helicobacter pylori determined by bacterial two-hybrid system

Abstract

The bacterium Helicobacter pylori infects more than 50% of the world population and causes several gastroduodenal diseases, including gastric cancer. Nevertheless, we still need to explore some protein interactions that may be involved in pathogenesis. MreB, an actin homolog, showed some special characteristics in previous studies, indicating that it could have different functions. Protein functions could be realized via protein-protein interactions. In the present study, the MreB protein from H. pylori 26695 fused with two tags 10×His and GST in tandem was overexpressed and purified from Escherchia coli. The purified recombinant protein was used to perform a pull-down assay with H. pylori 26695 cell lysate. The pulled-down proteins were identified by mass spectrometry (MALDI-TOF), in which the known important proteins related to morphogenesis were absent but several proteins related to pathogenesis process were observed. The bacterial two-hybrid system was further used to evaluate the protein interactions and showed that new interactions of MreB respectively with VacA, UreB, HydB, HylB and AddA were confirmed but the interaction MreB-MreC was not validated. These results indicated that the protein MreB in H. pylori has a distinct interactome, does not participate in cell morphogenesis via MreB-MreC but could be related to pathogenesis.

Biochemical and Biophysical Characterization of the Enolase fromHelicobacter pylori

“…From previous results from our lab, the expression of HpEno has been detected in the cell lysate of H. pylori 26695. Its presence was observed in the pull-down products using the protein MreB as a bait [22].…”

“…H. pylori has a reduced genome size (1.7 Mb), and many proteins have been lost in its evolutionary history, and the extant proteins can change their functions. In our laboratory, the enolase from H. pylori (HpEno) was pulled down with the protein MreB, an actin-like protein, indicating that HpEno can exist in a complex with MreB [22]. Thus, the enolase protein may have different structural and functional characteristics in this bacterium.…”

Morphology of Helicobacter pylori as a result of peptidoglycan and cytoskeleton rearrangements

“…In another study using the two-hybrid bacterial system, interaction between MreB and various H. pylori virulence factors was demonstrated, i.e. VacA (vacuolating cytotoxin; eukaryotic cells degradation), UreB (urease subunit; alkalisation of the acidic environment), HydB (hydrogenase subunit; maintenance of redox homeostasis), HylB (haemolysin secretion protein precursor; eukaryotic cells degradation), and AddA ( recB -like nuclease-helicase; DNA repair and recombination) [ 56 ]. What is more, the interaction of MreB with proteins involved in the H. pylori morphogenesis process was not observed.…”

Blue Light Sensing BlsA-Mediated Modulation of Meropenem Resistance and Biofilm Formation in Acinetobacter baumannii

“…In addition, the β-galactosidase activity of the tested strains harboring the complemented adenylate cyclase demonstrated the occurrence and degree of positive interactions under varied light and low temperature (23°C) conditions, suggesting that protein folding or interactions could also be affected by light and temperature ( Fig. 2C and D ) ( 34 , 35 ).…”