Detergent-Solubilized Patched Purified from Sf9 Cells Fails to Interact Strongly with Cognate Hedgehog or Ihog Homologs

patched;hedgehog;shh;hh;ptc;ihog;cdo;boc;Biophysics

Cleveland, Thomas EdgarAmzel, L. Mario ;

Johns Hopkins University

关键词: patched;    hedgehog;    shh;    hh;    ptc;    ihog;    cdo;    boc;    Biophysics;   

Others  :  https://jscholarship.library.jhu.edu/bitstream/handle/1774.2/39309/TEC%20Thesis%20Draft%202.docx?sequence=2&isAllowed=y

瑞士|英语

来源: JOHNS HOPKINS DSpace Repository

【 摘 要 】

The Hedgehog (Hh) signaling pathway mediates key tissue patterning events during animal development, and abnormal pathway activity is associated with several cancers. Hh proteins are secreted morphogens that specify cell fates in neighboring tissues in a concentration-dependent manner. The twelve-pass transmembrane protein Patched (Ptc) has been identified as a key Hh receptor in genetic and cell-based binding studies. In addition to Ptc, the CDO/Ihog family of co-receptors, and other accessory proteins, are necessary for proper Hh signaling. Structures of Hh proteins bound to members of the CDO/Ihog family are known, but the nature of the full Hh receptor complex is not well understood. We have expressed Ptc proteins from Drosophila and Mouse in Sf9 cells and find that purified, detergent-solubilized Ptc proteins do not interact strongly with cognate Hh and CDO/Ihog homologs. These results may reflect a nonnative conformation of purified Ptc or that an additional factor or factors is required for high-affinity Ptc binding to Hh.