1. Stoichiometry of TGF‐β receptor complexes determined by HPLC analysis.
Stoichiometry of TGF‐β receptor complexes determined by HPLC analysis. (A) Isolation of complexes of TGF‐β3 WW (solid line) and TGF‐β3 WD (dashed line) with the TβRI and TβRII extracellular domains using size exclusion chromotography. Small molecules that elute at the total volume are indicated by ‘SM’. (B) Peaks from panel (A) were analysed by SDS–PAGE gel under non‐reducing conditions; peaks a and b correspond to the complexes with TβRI and TβRII, whereas a’ and b’ correspond to excess unbound receptors. (C) Native gel analysis of the isolated TGF‐β3 WW and WD receptor complexes (left and right panels, respectively) either alone (lane 6) or with 2.0 molar equivalents of added TβRII (lane 7), TβRI (lane 8), or TβRI and TβRII (lane 9). (D) HPLC cation‐exchange analysis of the isolated TGF‐β3 WW and WD receptor complexes (solid and dashed lines, respectively) under denaturing conditions (8 M urea, pH 4.0). Identity of the eluted peaks is indicated.
Tao Huang et al. EMBO J. 2011;30:
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