TY - JOUR
T1 - Novel protein interactions with an actin homolog (MreB) of Helicobacter pylori determined by bacterial two-hybrid system
AU - Zepeda Gurrola, Reyna Cristina
AU - Fu, Yajuan
AU - Rodríguez Luna, Isabel Cristina
AU - Benítez Cardoza, Claudia Guadalupe
AU - López López, María de Jesús
AU - López Vidal, Yolanda
AU - Gutíerrez, Germán Rubén Aguilar
AU - Rodríguez Pérez, Mario A.
AU - Guo, Xianwu
N1 - Publisher Copyright:
© 2017 Elsevier GmbH
PY - 2017/8/1
Y1 - 2017/8/1
N2 - 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.
AB - 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.
KW - Actin homolog
KW - Bacterial two-hybrid system
KW - Helicobacter pylori
KW - Morphogenesis
KW - MreB
KW - Pathogenesis
KW - Pull-down assay
UR - http://www.scopus.com/inward/record.url?scp=85019144204&partnerID=8YFLogxK
U2 - 10.1016/j.micres.2017.04.008
DO - 10.1016/j.micres.2017.04.008
M3 - Artículo
C2 - 28602400
SN - 0944-5013
VL - 201
SP - 39
EP - 45
JO - Microbiological Research
JF - Microbiological Research
ER -