McGlynn Shawn E, Shepard Eric M, Winslow Mark A, Naumov Anatoli V, Duschene Kaitlin S, Posewitz Matthew C, Broderick William E, Broderick Joan B, Peters John W
Department of Chemistry and Biochemistry, The Astrobiology Biogeocatalysis Research Center, Montana State University, Bozeman, MT 59717, USA.
FEBS Lett. 2008 Jun 25;582(15):2183-7. doi: 10.1016/j.febslet.2008.04.063. Epub 2008 May 22.
In an effort to determine the specific protein component(s) responsible for in vitro activation of the [FeFe] hydrogenase (HydA), the individual maturation proteins HydE, HydF, and HydG from Clostridium acetobutylicum were purified from heterologous expressions in Escherichia coli. Our results demonstrate that HydF isolated from a strain expressing all three maturation proteins is sufficient to confer hydrogenase activity to purified inactive heterologously expressed HydA (expressed in the absence of HydE, HydF, and HydG). These results represent the first in vitro maturation of [FeFe] hydrogenase with purified proteins, and suggest that HydF functions as a scaffold upon which an H-cluster intermediate is synthesized.
为了确定负责[FeFe]氢化酶（HydA）体外激活的特定蛋白质成分，从丙酮丁醇梭菌中分离出的单个成熟蛋白HydE、HydF和HydG通过在大肠杆菌中的异源表达进行了纯化。我们的结果表明，从表达所有三种成熟蛋白的菌株中分离出的HydF足以赋予纯化的无活性异源表达HydA（在没有HydE、HydF和HydG的情况下表达）氢化酶活性。这些结果代表了首次使用纯化蛋白对[FeFe]氢化酶进行体外成熟，并表明HydF作为合成H簇中间体的支架发挥作用。
