Dimerization of translationally controlled tumor protein is essential for its cytokine-like activity.
BACKGROUND
Translationally Controlled Tumor Protein (TCTP) found in nasal lavage fluids of allergic patients was named IgE-dependent histamine-releasing factor (HRF). Human recombinant HRF (HrHRF) has been recently reported to be much less effective than HRF produced from activated mononuclear cells (HRFmn).


RESULTS
We found that only NH(2)-terminal truncated, but not C-terminal truncated, TCTP shows cytokine releasing activity compared to full-length TCTP. Interestingly, only NH(2)-terminal truncated TCTP, unlike full-length TCTP, forms dimers through intermolecular disulfide bonds. We tested the activity of dimerized full-length TCTP generated by fusing it to rabbit Fc region. The untruncated-full length protein (Fc-HrTCTP) was more active than HrTCTP in BEAS-2B cells, suggesting that dimerization of TCTP, rather than truncation, is essential for the activation of TCTP in allergic responses. We used confocal microscopy to evaluate the affinity of TCTPs to its putative receptor. We detected stronger fluorescence in the plasma membrane of BEAS-2B cells incubated with Del-N11TCTP than those incubated with rat recombinant TCTP (RrTCTP). Allergenic activity of Del-N11TCTP prompted us to see whether the NH(2)-terminal truncated TCTP can induce allergic airway inflammation in vivo. While RrTCTP had no influence on airway inflammation, Del-N11TCTP increased goblet cell hyperplasia in both lung and rhinal cavity. The dimerized protein was found in sera from allergic patients, and bronchoalveolar lavage fluids from airway inflamed mice.


CONCLUSIONS
Dimerization of TCTP seems to be essential for its cytokine-like activity. Our study has potential to enhance the understanding of pathogenesis of allergic disease and provide a target for allergic drug development.
Kim
Miyoung
M
College of Pharmacy, Center for Cell Signaling Research and Drug Discovery Research, Ewha Womans University, Seoul, Korea.
Min
Hyun Jung
HJ
Won
Hee Yeon
HY
Park
Heejin
H
Lee
Ji-Chul
JC
Park
Heung-Woo
HW
Chung
Junho
J
Hwang
Eun Sook
ES
Lee
Kyunglim
K
D000818:Animals; D001249:Asthma / Q000378:metabolism; D001483:Base Sequence; D014408:Biomarkers, Tumor / Q000502:physiology*; D001980:Bronchi / Q000166:cytology / Q000378:metabolism; D001992:Bronchoalveolar Lavage Fluid; D002460:Cell Line; D016207:Cytokines / Q000502:physiology*; D017931:DNA Primers; D019281:Dimerization; D004220:Disulfides / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D051379:Mice; D018613:Microscopy, Confocal; D000090324:Tumor Protein, Translationally-Controlled 1
D014408:Biomarkers, Tumor; D016207:Cytokines; D017931:DNA Primers; D004220:Disulfides; C000715647:TPT1 protein, human; C000715648:Tpt1 protein, mouse; C000715649:Tpt1 protein, rat; D000090324:Tumor Protein, Translationally-Controlled 1