Plant Physiol. Journal of Pharmacology and Experimental Therapeutics
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Published on July 9, 2008; 10.1104/pp.108.122846

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Two Arabidopsis thaliana ADP-glucose pyrophosphorylase large subunits (APL1 and APL2) are catalytic

Tiziana Ventriglia , Misty L. Kuhn , M Teresa Ruiz , Marina Ribeiro-Pedro , Federico Valverde , Miguel A. Ballicora , Jack Preiss , and Jose M. Romero *

Instituto de Bioquimica Vegetal y Fotosintesis, Centro de Investigaciones Cientificas Isla de la Cartuja, Universidad de Sevilla-C.S.I.C., c/ Americo Vespucio No 49, 41092-Sevilla, Spain, Department of Biochemistry, Michigan State University, East Lansing, Michigan 48824, and Dept. of Chemistry, Loyola University Chicago, 405 Flanner Hall, 1068 W. Sheridan Rd. Chicago, IL 60626

* Corresponding author; email: jmromero{at}ibvf.csic.es.

ADP-glucose pyrophosphorylase (ADP-Glc PPase) catalyzes the first committed step in starch biosynthesis. Higher plant ADP-Glc PPase is a heterotetramer ({alpha}2{beta}2) consisting of two small and two large subunits. There is increasing evidence that suggests that catalytic and regulatory properties of the enzyme from higher plants results from the synergy of both types of subunits. In Arabidopsis thaliana two genes encode small subunits (APS1 and APS2) and four large subunits (APL1 to 4). Here we show that in Arabidopsis thaliana APL1 and APL2, besides their regulatory role, have catalytic activity. Heterotetramers formed by combinations of a non-catalytic APS1 and the four large subunits showed that APL1 and APL2 exhibited ADP-Glc PPase activity with distinctive sensitivities to the allosteric activator (3-PGA). Mutation of the glucose-1-phosphate binding site of Arabidopsis and potato isoforms confirmed these observations. To determine the relevance of these activities in planta a T-DNA mutant of APS1 (aps1) was characterized. aps1 is starchless, lacked ADP-Glc PPase activity, APS1 mRNA, APS1 protein and is late flowering in long days. Transgenic lines of the aps1 mutant, expressing an inactivated form of APS1, recovered the wild type phenotype indicating that APL1 and APL2 have catalytic activity and may contribute to ADP-Glucose synthesis in planta.




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