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. 2004 Mar 16;322(1):93–104. doi: 10.1016/j.virol.2004.01.020

Fig. 5.

Fig. 5

Proteolytic analysis of Ad fibers. (A) Purified wild-type Ad41 long pentons LP41 (complex of penton base and long fiber), 0.2-μg portions. (B) Purified wild-type Ad41 short and long pentons in mixture, SP + LP41, 0.1-μg portions each. (C) Purified wild-type Ad5 pentons (P5, 1-μg portions) or Ad5 fiber (F5, 1-μg portions). The proteins were electrophoresed on a 7.5% or 10% SDS-PAGE gel at 4 °C. Lane 1, boiled fiber; lane 2, native fiber; lane 3, 1-min treatment with HCl, pH 2 at 37 °C; lane 4, fiber digested by chymotrypsin at pH 7.5 with ratio 1/5 enzyme/substrate; lane 5, fiber exposed to HCl for 1 min at pH 2 at 37 °C, neutralized, and digested by chymotrypsin at final pH 7.5 with ratio 1/5 enzyme/substrate. Fibers in pentons were revealed by Western blot with the appropriate antibody. Ad5 fiber was visualized by staining with Coomassie Brilliant Blue (F5, left side) or revealed by Western blot (F5, right side).