Determination of interactions between human thrombopoietin and its receptor MPL by yeast two-hybrid system and affinity biosensor.
The binding of human thrombopoietin to the extracellular domain of its receptor MPL prompts a cascade transduction of intracellular signals, leading to the development of megakaryocyte precursors and the production of circulating platelets. We have used a yeast two-hybrid system to reveal, via in vivo interactions between different deletion constructs of MPL and thrombopoietin, that the extracellular subunit 1 of MPL is the ligand binding site and the N-terminal domain of thrombopoietin alone is sufficient for the binding. The extracellular portion of MPL was heterologously expressed in E. coli and its specific affinity with thrombopoietin was visualized in vitro by resonance mirror biosensor technique.
Hsieh
D P
DP
Department of Biology, The Hong Kong University of Science and Technology, Clear Water Bay, Hong Kong, People's Republic of China.
Huxtable
S
S
Ng
K F
KF
Chen
H M
HM
Tsang
P W
PW
Wang
J
J
Xu
P P
PP
D001665:Binding Sites; D015374:Biosensing Techniques; D006801:Humans; D009363:Neoplasm Proteins; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D018121:Receptors, Cytokine; D053628:Receptors, Thrombopoietin; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D013926:Thrombopoietin / Q000235:genetics / Q000378:metabolism*; D013997:Time Factors; D020798:Two-Hybrid System Techniques; D015003:Yeasts / Q000235:genetics
D009363:Neoplasm Proteins; D011518:Proto-Oncogene Proteins; D018121:Receptors, Cytokine; D053628:Receptors, Thrombopoietin; D011994:Recombinant Proteins; C082218:MPL protein, human; D013926:Thrombopoietin