Immunocytochemical study of Bcr and Bcr-Abl localization in K562 cells.
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.
Telegeev
G D
GD
Institute of Molecular Biology and Genetics of NAS of Ukraine, Zabolotnogo 150, Kiev 03143, Ukraine.
Dubrovska
A N
AN
Nadgorna
V A
VA
Dybkov
M V
MV
Zavelevich
M P
MP
Maliuta
S S
SS
Gluzman
D F
DF
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
D000906:Antibodies; D011994:Recombinant Proteins; D016044:Fusion Proteins, bcr-abl; C496613:BCR protein, human; D051562:Proto-Oncogene Proteins c-bcr