This Article Full Text Full Text (PDF) All Versions of this Article: 136/2/3034    most recent pp.104.044040v1 Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in ISI Web of Science Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Citing Articles via ISI Web of Science (5) Citing Articles via Google Scholar Google Scholar Articles by Johansson, M. Articles by Knorpp, C. Search for Related Content PubMed PubMed Citation Articles by Johansson, M. Articles by Knorpp, C. Agricola Articles by Johansson, M. Articles by Knorpp, C.

BIOCHEMICAL PROCESSES AND MACROMOLECULAR STRUCTURES

Structure and Mutational Analysis of a Plant Mitochondrial Nucleoside Diphosphate Kinase. Identification of Residues Involved in Serine Phosphorylation and Oligomerization¹

Department of Plant Biology and Forest Genetics, Swedish University of Agricultural Sciences, S–750 07 Uppsala, Sweden (M.J., C.K.); and Department of Molecular Biology, Swedish University of Agricultural Sciences, S–751 24 Uppsala, Sweden (A.M.-H., I.A.)

We report the first crystal structure of a plant (Pisum sativum L. cv Oregon sugarpod) mitochondrial nucleoside diphosphate kinase. Similar to other eukaryotic nucleoside diphosphate kinases, the plant enzyme is a hexamer; the six monomers in the asymmetric unit are arranged as trimers of dimers. Different functions of the kinase have been correlated with the oligomeric structure and the phosphorylation of Ser residues. We show that the occurrence of Ser autophosphorylation depends on enzymatic activity. The mutation of the strictly conserved Ser-119 to Ala reduced the Ser phosphorylation to about one-half of that observed in wild type with only a modest change of enzyme activity. We also show that mutating another strictly conserved Ser, Ser-69, to Ala reduces the enzyme activity to 6% and 14% of wild-type using dCDP and dTDP as acceptors, respectively. Changes in the oligomerization pattern of the S69A mutant were observed by cross-linking experiments. A reduction in trimer formation and a change in the dimer interaction could be detected with a concomitant increase of tetramers. We conclude that the S69 mutant is involved in the stabilization of the oligomeric state of this plant nucleoside diphosphate kinase.

² These authors contributed equally to the paper.

Article, publication date, and citation information can be found at www.plantphysiol.org/cgi/doi/10.1104/pp.104.044040.

^* Corresponding author; e-mail carina.knorpp{at}vbsg.slu.se; fax 46–18–67–32–79.

Received April 2, 2004; returned for revision July 21, 2004; accepted August 5, 2004.

This article has been cited by other articles:

				S. Dorion, F. Dumas, and J. Rivoal Autophosphorylation of Solanum chacoense cytosolic nucleoside diphosphate kinase on Ser117 J. Exp. Bot., December 1, 2006; 57(15): 4079 - 4088. [Abstract] [Full Text] [PDF]

				J.-D. Pedelacq, G. S. Waldo, S. Cabantous, E. C. Liong, and T. C. Terwilliger Structural and functional features of an NDP kinase from the hyperthermophile crenarchaeon Pyrobaculum aerophilum Protein Sci., October 1, 2005; 14(10): 2562 - 2573. [Abstract] [Full Text] [PDF]