title: Immunocytochemical study of Bcr and Bcr-Abl localization in K562 cells.
abstract: OBJECTIVE
  To obtain polyclonal antibodies against recombinant proteins recognizing Bcr domain and fusion region of Bcr-Abl and analyze the patterns of intracellular distribution of Bcr and Bcr-Abl proteins in K562 cells of chronic myelogenous leukemia.
  
  
  METHODS
  The coding sequences of DH and PH domains of Bcr-Abl were cloned, and the recombinant proteins were expressed in E. coli. The rabbit polyclonal antibodies were produced and used for immunocytochemical study of Bcr and Bcr-Abl localization in K562 cells.
  
  
  RESULTS
  The gene constructs containing sequences coding for DH and PH domains of Bcr-Abl have been obtained. The antibodies with relative specificity to corresponding recombinant proteins differ by the patterns of their intracellular reactivity with Bcr- and Bcr-Abl related structures. While Bcr protein is located predominantly perinuclearly, antibody against hybrid Bcr-Abl protein is reacted with the structures in cell periphery, namely on cell membranes.
  
  
  CONCLUSIONS
  Antibodies against DH and PH domains of Bcr-Abl react with proteins located differently in chronic myelogenous leukemia cells. The difference in intracellular localization of Bcr and Bcr-Abl may be attributable to the different domains interacting with different multiprotein complexes.
authors.affiliation: Institute of Molecular Biology and Genetics of NAS of Ukraine, Zabolotnogo 150, Kiev 03143, Ukraine.
authors.forename: G D
authors.initials: GD
authors.lastname: Telegeev
authors.forename: A N
authors.initials: AN
authors.lastname: Dubrovska
authors.forename: V A
authors.initials: VA
authors.lastname: Nadgorna
authors.forename: M V
authors.initials: MV
authors.lastname: Dybkov
authors.forename: M P
authors.initials: MP
authors.lastname: Zavelevich
authors.forename: S S
authors.initials: SS
authors.lastname: Maliuta
authors.forename: D F
authors.initials: DF
authors.lastname: Gluzman
mesh_terms: D000818:Animals; D000906:Antibodies; D000918:Antibody Specificity; D015153:Blotting, Western; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D016044:Fusion Proteins, bcr-abl / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000235:genetics / Q000378:metabolism*; D009369:Neoplasms; D017434:Protein Structure, Tertiary; D051562:Proto-Oncogene Proteins c-bcr / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D011817:Rabbits; D011994:Recombinant Proteins / Q000378:metabolism
chemical_list: D000906:Antibodies; D011994:Recombinant Proteins; D016044:Fusion Proteins, bcr-abl; C496613:BCR protein, human; D051562:Proto-Oncogene Proteins c-bcr