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. 1998 Jan;9(1):117–130. doi: 10.1091/mbc.9.1.117

Figure 6.

Figure 6

Signal sequence-independent binding of RNCs to PK-RM. Ribosome-ffLuc77 complexes were assembled in a reticulocyte lysate system. (A) Aliquots of (a) high-salt extracted RNCs (-NAC), (b) low-salt extracted RNCs (+NAC) and (c, d) total translation products (c, 2.5 μl; d, 5 μl) were incubated with 8 eq of PK-RM. The RNCs (a and b) were derived from 10 μl of total translation products. Membrane-bound ffLuc77 was separated from unbound ffLuc77 by flotation on discontinuous sucrose gradients as in Figure 1A. (B) Total ffLuc77 translation products were incubated with 2 eq of PK-RM (a and b) or 4 eq of PK-RM (c and d) before flotation on discontinuous sucrose gradients. Aliquots of the three fractions from the gradients were precipitated with either 67% saturated ammonium sulfate (a and c) or with CTABr (b and d) before PAGE in SDS. The percent of ffLuc77 that was recovered with the microsomes was calculated as described in MATERIALS AND METHODS.