Search academic texts

Information × Registration Number 0209U002337, 0104U000163 , R & D reports Title Studying of internal factors of tumour progression in human malignant lymphomes and myeloproliferative diseases. popup.stage_title Head Мaliuta Stanislav Staniclavovych, Telegeev Gennadiy Dmytrovych, Registration Date 09-02-2009 Organization Institute of molecular biology & genetics popup.description2 The aim of the work was studying the functional features of Bcr DH and PH domains and. For that purpose we cloned PH, DH and DHPH part of bcr gene and subcloned it into bacterial expression vector pET32a.Recombinant his-tagged PH protein was purified on Ni-NTA column from BL21 DE3 E.Coli strain cell lysates. CD spectra of recombinant PH protein were analyzed and secondary structure content was identified.Experimental data suggest that PH domain binds to phosphatidylinositol monophosphates PI(3)P, PI(4)P and PI(5)P with high affinity. There is uneven lipid content in intracellular cell membranes. Therefore, PH domain binding to phosphainositols might effect p210 Bcr-Abl protein localization. This was confirmed by comparison of p190 and p210 Bcr-Abl cell localization in immunofluorescent microscopy assay. Protein-protein interactions of PH domain were studied by pull-down assay.Peptide spectra were collected by MALDI TOF mass-spectrometry and corresponding proteins were identified by ProFound web resource. We identified 23 proteins that potentially bind to Bcr PH domain.GEF activity of Bcr DH domain towards RhoA, Rac1 and Cdc42 hasn't been detected in both in vitro and in vivo experiments. Taken together, the study reveals important functions of Bcr DH and PH domains that could affect Bcr-Abl signal pathways.Expression of differet genes of DNMT for patients with myeloproliferative diseases wan detected. Product Description popup.authors popup.nrat_date 2020-04-02 Close