3UIE

Crystal structure of adenosine 5'-phosphosulfate kinase from Arabidopsis Thaliana in Complex with AMPPNP and APS

Summary for 3UIE

• Entry DOI: 10.2210/pdb3uie/pdb
• Descriptor: Adenylyl-sulfate kinase 1, chloroplastic, ADENOSINE-5'-PHOSPHOSULFATE, PHOSPHOAMINOPHOSPHONIC ACID-ADENYLATE ESTER, ... (5 entities in total)
• Functional Keywords: rossmann fold, kinase, chloroplast, transferase-transferase inhibitor complex, transferase/transferase inhibitor
• Biological source: Arabidopsis thaliana (mouse-ear cress,thale-cress)
• Cellular location: Plastid, chloroplast : Q43295
• Total number of polymer chains: 3
• Total formula weight: 69186.31

Authors

Ravilious, G.E.,Jez, J.M. (deposition date: 2011-11-04, release date: 2012-01-25, Last modification date: 2017-11-08)

Primary citation

Ravilious, G.E.,Nguyen, A.,Francois, J.A.,Jez, J.M.
Structural basis and evolution of redox regulation in plant adenosine-5'-phosphosulfate kinase.
Proc.Natl.Acad.Sci.USA, 109:309-314, 2012

PubMed Abstract: Adenosine-5'-phosphosulfate (APS) kinase (APSK) catalyzes the phosphorylation of APS to 3'-phospho-APS (PAPS). In Arabidopsis thaliana, APSK is essential for reproductive viability and competes with APS reductase to partition sulfate between the primary and secondary branches of the sulfur assimilatory pathway; however, the biochemical regulation of APSK is poorly understood. The 1.8-Å resolution crystal structure of APSR from A. thaliana (AtAPSK) in complex with β,γ-imidoadenosine-5'-triphosphate, Mg(2+), and APS provides a view of the Michaelis complex for this enzyme and reveals the presence of an intersubunit disulfide bond between Cys86 and Cys119. Functional analysis of AtAPSK demonstrates that reduction of Cys86-Cys119 resulted in a 17-fold higher k(cat)/K(m) and a 15-fold increase in K(i) for substrate inhibition by APS compared with the oxidized enzyme. The C86A/C119A mutant was kinetically similar to the reduced WT enzyme. Gel- and activity-based titrations indicate that the midpoint potential of the disulfide in AtAPSK is comparable to that observed in APS reductase. Both cysteines are invariant among the APSK from plants, but not other organisms, which suggests redox-control as a unique regulatory feature of the plant APSK. Based on structural, functional, and sequence analyses, we propose that the redox-sensitive APSK evolved after bifurcation of the sulfur assimilatory pathway in the green plant lineage and that changes in redox environment resulting from oxidative stresses may affect partitioning of APS into the primary and secondary thiol metabolic routes by having opposing effects on APSK and APS reductase in plants.

PubMed: 22184237
DOI: 10.1073/pnas.1115772108

Experimental method

X-RAY DIFFRACTION (1.794 Å)