Association of in vitro translated amino-terminal moesin with PSGL-1 and ICAM-3.

In vitro translated amino-terminal moesin (N-moesin) was incubated with GST or GST-fusion proteins containing the cytoplasmic domain of PSGL-1 or ICAM-3 linked to glutathione-Sepharose beads. After incubation, Sepharose beads were centrifuged and washed and bound proteins were boiled in sample buffer and analyzed by means of 10% SDS-PAGE and autoradiography. N-moesin labeled with sulfur 35 was loaded as a control in the last lane. Autoradiography showed nonspecific binding of a polypeptide of about 50 kd and specific binding of a labeled polypeptide of the size of N-moesin to PSGL-1 and ICAM-3. Molecular masses in kilodaltons are indicated on the left.