Ferrero Danisa M L, Asencion Diez Matias D, Kuhn Misty L, Falaschetti Christine A, Piattoni Claudia V, Iglesias Alberto A, Ballicora Miguel A
Laboratorio de Enzimología Molecular, Instituto de Agrobiotecnología del Litoral (CONICET - UNL), Facultad de Bioquímica y Ciencias Biológicas, Universidad Nacional del Litoral, Santa Fe, Argentina.
Department of Chemistry and Biochemistry, Loyola University Chicago, Chicago, IL, United States.
Front Plant Sci. 2018 Oct 16;9:1498. doi: 10.3389/fpls.2018.01498. eCollection 2018.
The ADP-glucose pyrophosphorylase from wheat endosperm controls starch synthesis in seeds and has unique regulatory properties compared to others from this family. It comprises two types of subunits, but despite its importance little is known about their roles. Here, we synthesized the wheat endosperm ADP-glucose pyrophosphorylase small (S) and large (L) subunit genes, heterologously expressed them in , and kinetically characterized the recombinant proteins. To understand their distinct roles, we co-expressed them with well characterized subunits from the potato tuber enzyme to obtain hybrids with one S subunit from one source and an L subunit from the other. After kinetic analyses of these hybrids, we concluded that the unusual insensitivity to activation of the wheat endosperm enzyme is caused by a pre-activation of the L subunit. In addition, the heat stability and sensitivity to phosphate are given by the S subunit.
来自小麦胚乳的ADP - 葡萄糖焦磷酸化酶控制种子中的淀粉合成,与该家族的其他酶相比具有独特的调节特性。它由两种类型的亚基组成,但尽管其很重要,人们对它们的作用却知之甚少。在这里,我们合成了小麦胚乳ADP - 葡萄糖焦磷酸化酶小(S)亚基和大(L)亚基基因,在体外进行异源表达,并对重组蛋白进行动力学表征。为了解它们各自的作用,我们将它们与来自马铃薯块茎酶的已充分表征的亚基共表达,以获得具有来自一个来源的一个S亚基和来自另一个来源的一个L亚基的杂合体。对这些杂合体进行动力学分析后,我们得出结论,小麦胚乳酶对激活的异常不敏感性是由L亚基的预激活引起的。此外,热稳定性和对磷酸盐的敏感性由S亚基决定。