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. 2010 Sep 27;285(48):37263–37270. doi: 10.1074/jbc.M110.170027

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© 2010 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 2. — Ccc2 and RAN1 copper transporters and saturable ethylene binding to ETR1 expressed in yeast. A, intact yeast cells expressing the ETR1 receptor in the Ccc2 (Ccc2-ETR1) or Δccc2 (Δccc2-ETR1) background were analyzed for [¹⁴C]ethylene binding. The effects of transforming Δccc2-ETR1 yeast with RAN1, ran1-1, and ran1-2 on ETR1 ethylene binding are shown. Ethylene binding using [¹⁴C]ethylene is compared between samples treated with [¹⁴C]ethylene (0.1 μl/liter) and identical samples treated with [¹⁴C]ethylene (0.1 μl/liter) plus [¹²C]ethylene (100 μl/liter). Equal amounts of yeast used in the binding assays were analyzed on Western blots probed with anti-ETR1 antibodies. B, displaceable ethylene binding in the presence and absence of 300 μm copper sulfate is shown for membranes isolated from Ccc2-ETR1 and Δccc2-ETR1 yeast. Displaceable binding was calculated by subtracting the amount of [¹⁴C]ethylene (0.1 μl/liter) bound in the presence of excess [¹²C]ethylene (100 μl/liter) from amount of [¹⁴C]ethylene (0.1 μl/liter) bound in the absence of added [¹²C]ethylene. In both panels, the mean ± S.D. for disintegrations/min/g of yeast is shown.