Solubilization and some Properties of the Erythrocyte Receptor for Adenovirus Type 7 Haemagglutinin

Abstract

WE describe here the solubilization and some of the properties of the erythrocyte receptor for the haemagglutinin (HA) of adenovirus type 7, a representative of Rosen's subgroup 1 of adenoviruses¹. Ghosts prepared from rhesus monkey red blood cells (RBC) as described by Marchesi and Palade² were suspended in 0.05 M tris HCl buffer (pH 7.0), and treated for 1 h at 37° C with the enzymes listed in Table 1. The action of trypsin was stopped by the addition of soybean trypsin inhibitor (0.05 mg/ml.). Ghosts treated with α-chymotrypsin, papain or pronase were pelleted by centrifugation at 45,000g for 10 min, resuspended in the original volume in phosphate buffered saline (PBS; 0.075 M phosphate, 0.072 M NaCl, pH 7.2) and titrated for receptor activity. Ficin treatment was performed in the presence of cysteine (1 mg/ml.) and carboxypeptidase treatment in the presence of MgSO₄ (0.001 M). At the end of the treatment, disodium ethylenediaminetetraacetate was added up to final concentrations of 0.003 and 0.005 M respectively.