RESUMO
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.
Assuntos
Actinas/metabolismo , Proteínas de Bactérias/metabolismo , Helicobacter pylori/metabolismo , Domínios e Motivos de Interação entre Proteínas/fisiologia , Técnicas do Sistema de Duplo-Híbrido , Actinas/fisiologia , Proteínas de Bactérias/química , Proteínas de Bactérias/genética , Proteínas de Bactérias/fisiologia , DNA Bacteriano/genética , Escherichia coli/genética , Regulação Bacteriana da Expressão Gênica , Genes Bacterianos , Helicobacter pylori/genética , Espectrometria de Massas , Proteínas de Membrana/metabolismo , Proteínas de Membrana/fisiologia , Morfogênese , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismoRESUMO
Hp diagnostic is made by invasive methods using gastric biopsy of antrum, for culture and histological study. Non invasive are serology and urea breath test. 152 Hp from 19 children's with acute gastritis (46. 1%); 9 Hp from an adult. There had ampicillin resistance, 27. 4%, claritromicin 21. 8%, metronidazol 58. 4% and tetracycline 31. 5%, the 21 % susceptible to four anti-microbial. RAPD-PCR with 4 primers gave 44 profiles, related with clinical profile. In 7 children, there were two profiles RAPD. Three were similar but different each other. AFLP 23 adults 151 Hp; cagA and vacA genotypes gave unique patterns for each patient. The genes cagA, babA and oipA, secuencing and compared with reported Hp (Genbank); gave high polymorfism. Everything indicates that there are colonization with multiple Hp strains in Mexicans. In 645 sera from 352 children: 36.9% with Hp, the 46. 9% gave him/her anti-shits and 16. 2% antibodies to urease. Adults 293 (89. 1%) with Hp, 78. 9% gave anti-CagA and 59% antibodies to urease. The expression of IL-8 was bigger in infected children that in not infected and more significant in peptic ulcer. Genomic library Cag-PAI revealed 90% (Hp) positive, they had the complete PAI, 2 were incomplete and three negatives. PCR -LiPA LiPA (LineProbe Assay) is suggestive for genotyping assays.
