Abstract
NADP-dependent chloroplastic malate dehydrogenase (E.C.1.1.1.82) is regulated by thiol disulfide-interchange with thioredoxin. It displays two regulatory disulfides per subunit, located in specific sequence extensions respectively at the N- and C-terminal ends of each subunit. In the present study, attempts were made to transfer the regulatory properties of sorghum NADP-malate dehydrogenase to a constitutively active NAD-dependent malate dehydogenase (E.C.1.1.1.37) from the thermophilic bacteria Thermus flavus, by grafting the regulatory extensions of the former to the latter. The results demonstrate that a successful transfer of redox regulation properties requires the grafting of both full-length extensions, but also the introduction of specific hydrophobic residues in the core part of the protein. These residues are very likely involved in the interaction between monomers, and structural changes at the active site.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- MDH:
-
Malate dehydrogenase
- PCR:
-
Polymerase chain reaction
- TRX:
-
Thioredoxin
- WT:
-
Wild-type
References
Amman E, Ochs B, Abel K-J (1988) Tightly regulated tac promoter vectors useful for the expression of unfused and fused proteins in Escherichia coli. Gene 69:301–315
Bald D, Noji H, Stumpp MT, Yoshida M, Hisabori T (2000) ATPase activity of a highly stable α3β3γ subcomplex of thermophilic F1 can be regulated by the introduced regulatory region of γ subunit of chloroplast F1. J Biol Chem 275:12757–12762
Carr P, Verger D, Ashton AR, Ollis D (1999) Chloroplast NADP-malate dehydrogenase: structural basis of light-dependent regulation of activity by thiol oxidation and reduction. Structure 7:461–475
Cazalis R, Chueca A, Sahrawy M, Lopez-Gorge J (2004) Construction of chimeric cytosolic fructose-1,6-bisphosphatases by insertion of a chloroplastic redox regulatory cluster. J Physiol Biochem 60:7–21
de Lamotte-Guery F, Miginiac-Maslow M, Decottignies P, Stein M, Minard P, Jacquot JP (1991) Mutation of a negatively charged amino acid in thioredoxin modifies its reactivity with chloroplastic enzymes. Eur J Biochem 196:287–294
Goyer A, Decottignies P, Issakidis-Bourguet E, Miginiac-Maslow M (2001) Sites of interaction of thioredoxin with sorghum NADP-malate dehydrogenase. FEBS Lett 505:405–408
Hatch MD, Agostino A (1992) Bi-level disulfide group reduction in the activation of C4 leaf nicotinamide adenine dinucleotide phosphate malate dehydrogenase. Plant Physiol 100:360–366
Hirasawa M, Ruelland E, Schepens I, Issakidis-Bourguet E, Miginiac-Maslow M, Knaff DB (2000) Oxidation–reduction properties of the regulatory disulfides of sorghum chloroplast NADP-malate dehydrogenase. Biochemistry 39:3344–3350
Iijima S, Uozumi T, Beppu T (1986) Molecular cloning of Thermus flavus malate dehydrogenase gene. Agric Biol Chem 50:589–592
Issakidis E, Miginiac-Maslow M, Decottignies P, Jacquot JP, Crétin C, Gadal P (1992) Site-directed mutagenesis reveals the involvement of an additional thioredoxin-dependent regulatory site in the activation of recombinant sorghum leaf NADP-malate dehydrogenase. J Biol Chem 267:21577–21583
Issakidis E, Saarinen M, Decottignies P, Jacquot JP, Crétin C, Gadal P, Miginiac-Maslow M (1994) Identification and characterization of the second regulatory disulfide bridge of recombinant sorghum leaf NADP-malate dehydrogenase. J Biol Chem 269:3511–3517
Johansson K, Ramaswamy S, Saarinen M, Lemaire-Chamley M, Issakidis-Bourguet E, Miginiac-Maslow M, Eklund H (1999) Structural basis for light activation of a chloroplast enzyme. The structure of sorghum NADP-malate dehydrogenase in its oxidized form. Biochemistry 38:4319–4326
Kelly CA, Nishiyama M, Ohnishi Y, Beppu T, Birktoft JJ (1993) Determinants of protein thermostability observed in the 1.9-Å crystal structure of malate dehydrogenase from the thermophilic bacterium Thermus flavus. Biochemistry 32:3913–3922
Koradi R, Billeter M, Wüthrich K (1996) MOLMOL: a program for display and analysis of macromolecular structures. J Mol Graph 14:51–55
Krenn BE, Strotmann H, Van Walraven HS, Scholts MJC, Kraayenhof R (1997) The ATP synthase γ subunit provides the primary site of activation of the chloroplast enzyme: experiments with a chloroplast-like Synechocystis 6803 mutant. Biochem J 323:841–845
Krimm I, Goyer A, Issakidis-Bourguet E, Miginiac-Maslow M, Lancelin JM (1999) Direct NMR observation of the thioredoxin-mediated reduction of the chloroplast NADP-malate dehydogenase provides a structural basis for the relief of auto-inhibition. J Biol Chem 274:34539–34542
Lemaire SD, Quesada A, Merchan F, Corral JM, Igeno MI, Keryer E, Issakidis-Bourguet E, Hirasawa M, Knaff DB, Miginiac-Maslow M (2005) NADP-malate dehydrogenase from the unicellular green alga Chlamydomonas reinhardtii: a first step towards redox regulation? Plant Physiol 137:514–521
Miginiac-Maslow M, Lancelin JM (2002) Intrasteric inhibition in redox signaling: light activation of NADP-malate dehydrogenase. Photosyn Res 72:1–12
Nishiyama M, Matsubara N, Yamamoto K, Iijima S, Uozumi T, Beppu T (1986) Nucleotide sequence of the malate dehydrogenase gene of Thermus flavus and its mutation directing an increase in enzyme activity. J Biol Chem 261:14178–14183
Ruelland E, Miginiac-Maslow M (1999) Regulation of chloroplast enzyme activities by thiol-disulfide interchange with reduced thioredoxin: activation or relief from inhibition? Trends Plant Sci 4:136–141
Ruelland E, Lemaire-Chamley M, Le Maréchal P, Issakidis-Bourguet E, Djukic N, Miginiac-Maslow M (1997) An internal cysteine is involved in the thioredoxin-dependent activation of sorghum NADP-malate dehydrogenase. J Biol Chem 272:19851–19857
Ruelland E, Johansson K, Decottignies P, Djukic N, Miginiac-Maslow M (1998) The auto-inhibition of sorghum NADP-malate dehydrogenase is mediated by a C-terminal negative charge. J Biol Chem 273:33482–33488
Sali A, Potterton L, Yuan F, van Hijmen H, Karplus M (1995) Evaluation of comparative protein modelling by MODELLER. Proteins 23:318–326
Schepens I, Johansson K, Decottignies P, Gillibert M, Hirasawa M, Knaff DB, Miginiac-Maslow M (2000a) Inhibition of the thioredoxin-dependent activation of the NADP-malate dehydrogenase and cofactor specificity. J Biol Chem 275:20996–21001
Schepens I, Ruelland E, Miginiac-Maslow M, Le Maréchal P, Decottignies P (2000b) The role of active site arginines of sorghum NADP-malate dehydrogenase in thioredoxin-dependent activation and activity. J Biol Chem 275:35792–35798
Schürmann P, Jacquot JP (2000) Plant thioredoxin systems revisited. Annu Rev Plant Physiol Plant Mol Biol 51:371–400
Werner-Grüne S, Gunkel D, Schumann J, Strotmann H (1994) Insertion of a “chloroplast-like” regulatory segment responsible for thiol modulation into gamma-subunit of F0F1-ATPase of the cyanobacterium Synechocystis 6803 by mutagenesis of atpC. Mol Gen Genet 244:144–150
Acknowledgments
We are grateful to E. Keryer for skillful technical assistance and to Prof. T. Beppu and Dr. M. Nishiyama (Tokyo University) for the gift of the T. flavus NAD-MDH clone and the mdh-E. coli strain.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Issakidis-Bourguet, E., Lavergne, D., Trivelli, X. et al. Transferring redox regulation properties from sorghum NADP-malate dehydrogenase to Thermus NAD-malate dehydrogenase. Photosynth Res 89, 213–223 (2006). https://doi.org/10.1007/s11120-006-9094-4
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11120-006-9094-4