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. Author manuscript; available in PMC: 2010 Apr 29.
Published in final edited form as: Chem Biol. 2008 Nov 24;15(11):1175–1186. doi: 10.1016/j.chembiol.2008.09.013

Figure 4. Substrate preference of BryP constructs.

Figure 4

(A) Autoradiography of SDS-PAGE of BryP AT1 substrate preference experiment. Lanes 1 & 2 = malonyl-CoA/[14C]-malonyl-CoA, lanes 3 & 4 = methylmalonyl-CoA/[14C]-malonyl-CoA, lanes 5 & 6 = methylmalonyl-CoA/[14C]-methylmalonyl-CoA, and lanes 7 & 8 = malonyl-CoA/[14C]-methylmalonyl-CoA. Lanes 1, 3, 5, 7 contain BryP AT1 only, and lanes 2, 4, 6, 8 contain BryP AT1 and BryB M7 ACP. (B) Autoradiography of SDS-PAGE of BryP constructs loading onto BryB M7 ACP. Lanes 1 & 6 = BryP AT1 + BryB M7 ACP, lanes 2 & 7 = BryP AT1AT2 + BryB M7 ACP, lanes 3 & 8 = BryP AT2(32) + BryB M7 ACP, lanes 4 & 7 = BryP AT2(37) + BryB M7 ACP, lanes 5 & 10 = BryP AT2(47) + BryB M7 ACP, and lane L = ladder. Lanes 1 – 5 = [14C]-malonyl-CoA and lanes 6–10 = [14C]-methylmalonyl-CoA. (C) Kinetics of BryP AT1 loading [14C]-malonyl-CoA onto loading onto BryB M7 ACP. Reactions and no enzyme controls were run in triplicate. (D) FT-ICR MS analysis (+37 charge state) of full BryP AT1 incubated with mixture of acyl-CoAs (top panels) or no substrate (bottom panels). Asterisk indicates peak that is magnified (right panels).