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. 2005 Jan;16(1):238–247. doi: 10.1091/mbc.E04-06-0534

Figure 4.

Figure 4.

Fusion of GFP to the N terminus of PakB results in intracellular proteolysis. (A) Immunoblot analysis using an anti-PakB antibody of PakB-null cells expressing PakB constructs. Extracts were obtained from cells expressing PakB with an N-terminal 7 × His tag (His-PakB), PakB with GFP fused to the N terminus (4 clones labeled GFP-PakB1-4), PakB-ΔN with GFP fused to the N terminus (GFP-PakB-ΔN), PakB-ΔPL with GFP fused to the N terminus (GFP-PakB-ΔPL), and PakB with GFP fused to the C terminus (PakB-GFP). GFP-PakB, expected to have a size of 140 kDa (arrow), is degraded to products of 110 and 95 kDa in cell lines GFPPakB1-3. The other PakB fusion proteins exhibit electrophoretic mobilities consistent with their predicted molecular masses. (B) Immunoblot analysis using an anti-GFP antibody shows that the 110- and 95-kDa GFP-PakB degradation products retain an intact N terminus. For each lane in A and B, 20 μg of protein was loaded. Molecular mass standards are indicated in kilodaltons.