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. 2014 Sep 3;27(10):1757–1768. doi: 10.1021/tx500229h

Figure 6.

Figure 6

Secosterol dehydration products and their reactivity. (top scheme) Secosterols and their dehydration products. MALDI-TOF-MS analysis of cyt c modification with secosterol dehydration products. (A) Modification with seco A–H2O. (B) Modification with seco B–H2O (II). (C) Modification with seco B–H2O (I). (D) Modification with seco B–2H2O. (E) Streptavidin fluorophore visualization of HSA modification with a-seco A compared to a-seco A–H2O. [The higher weight protein (∼130 kDa) is HSA dimer.] (F) Integrated intensities from streptavidin fluorophore analysis of HSA (1 mg/mL) modification with a-seco A (20 μM) compared to a-seco A–H2O (20 μM). Alkynyl sterol-adducted protein was tagged with biotin via click reaction and labeled with the streptavidin–AlexaFluor 680 conjugate. The fluorophore was detected and integrated intensities were obtained using the Odyssey Infrared Imaging System and Application Software. For each streptavidin fluorophore analysis, the lane with the lowest integrated intensity was set to 0.