Abstract
The brown alga Laminaria digitata features a distinct vanadium-dependent iodoperoxidase (vIPO) activity, which has been purified to electrophoretic homogeneity. Steady-state analyses at pH 6.2 are reported for vIPO (K I−m =2.5 mM; k I−cat =462 s−1) and for the previously characterised vanadium-dependent bromoperoxidase in L. digitata (K I−m =18.1 mM; k I−cat =38 s−1). Although the vIPO enzyme specifically oxidises iodide, competition experiments with halides indicate that bromide is a competitive inhibitor with respect to the fixation of iodide. A full-length complementary ANA (cDNA) was cloned and shown to be actively transcribed in L. digitata and to encode the vIPO enzyme. Mass spectrometry analyses of tryptic digests of vIPO indicated the presence of at least two very similar proteins, in agreement with Southern analyses showing that vIPOs are encoded by a multigenic family in L. digitata. Phylogenetic analyses indicated that vIPO shares a close common ancestor with brown algal vanadium-dependent bromoperoxidases. Based on a three-dimensional structure model of the vIPO active site and on comparisons with those of other vanadium-dependent haloperoxidases, we propose a hypothesis to explain the evolution of strict specificity for iodide in L. digitata vIPO.
Abbreviations
- bp:
-
base pair
- BPO:
-
Bromoperoxidase
- cDNA:
-
Complementary DNA
- CPO:
-
Chloroperoxidase
- HPO:
-
Haloperoxidase
- IPO:
-
Iodoperoxidase
- LC:
-
Liquid chromatography
- mRNA:
-
Messenger RNA
- MS/MS:
-
Tandem mass spectrometry
- PAGE:
-
Polyacrylamide gel electrophoresis
- PCR:
-
Polymerase chain reaction
- RACE:
-
Rapid amplification of cDNA ends
- SDS:
-
Sodium dodecyl sulphate
- SSC:
-
Saline–sodium citrate
- UTR:
-
Untranslated region
- vBPO:
-
Vanadium-dependent BPO
- vCPO:
-
Vanadium-dependent CPO
- vHPO:
-
Vanadium-dependent HPO
- vIPO:
-
Vanadium-dependent IPO
References
Gribble GW (2003) Chemosphere 52:289–297
Littlechild JA (1999) Curr Opin Chem Biol 3:28–34
de Boer E, Wever R (1988) J Biol Chem 263:12326–12332
Arber JM, de Boer E, Garner CD, Hasnain SS, Wever R (1989) Biochemistry 28:7968–7973
Vilter H (1995) In: Sigel H, Sigel A (eds) Metal ions in biological systems. vol. 31 Marcel Dekker, Inc., New York, Basel, Hong Kong, pp325–362
Vollenbroek EGM, Simons LH, van Schijndel JW, Barnett P, Balzar M, Dekker HL, van der Linden C, Wever R (1995) Biochem Soc Trans 23:267–271
Barnett P, Hemrika W, Dekker HL, Muijsers AO, Renirie R, Wever R (1998) J Biol Chem 273:23381–23387
Plat H, Krenn BE, Wever R (1987) Biochem J 248:277–279
Messerschmidt A, Wever R (1996) Proc Natl Acad Sci USA 93:392–396
Weyand M, Hecht HJ, Kiess M, Liaud M-F, Vilter H, Schomburg D (1999) J Mol Biol 293:595–611
Isupov MN, Dalby AR, Brindley AA, Izumi Y, Tanabe T, Murshudov GN, Littlechild JA (2000) J Mol Biol 299:1035–1049
Butler A, Carter JN, Simpson MT (2001) In: Bertini I, Sigel A, Sigel H (eds) Handbook on metalloproteins. Marcel Dekker, Inc, New York, Basel uijsers AO, pp153–179
Tanaka N, Hasan Z, Wever R (2003) Inorg Chim Acta 356:288–296
Carter JN, Beatty KE, Simpson MT, Butler A (2002) J Inorg Biochem 91:59–69
Messerschmidt A, Prade L, Wever R (1997) Biol Chem 378:309–315
Hemrika W, Renirie R, Macedo-Ribeiro S, Messerschmidt A, Wever R (1999) J Biol Chem 274:23820–23827
Butler A (1999) Coord Chem Rev 187:17–35
Renirie R, Hemrika W, Wever R (2000) J Biol Chem 275:11650–11657
Dau H, Dittmer J, Epple M, Hanss J, Kiss E, Rehder D, Schulzke C, Vilter H (1999) FEBS Lett 457:237–240
Colin C, Leblanc C, Wagner E, Delage L, Leize-Wagner E, van Dorsselaer A, Kloareg B, Potin P (2003) J Biol Chem 278:23545–23552
Apt KE, Clendennen SK, Powers DA, Grossman AR (1995) Mol Gen Genet 246:455–464
Crépineau F, Roscoe T, Kaas R, Kloareg B, Boyen C (2000) Plant Mol Biol 43:503–513
Katoh K, Misawa K, Kuma K, Miyata T (2002) Nucleic Acids Res 30:3059–3066
Gouet P, Robert X, Courcelle E (2003) Nucleic Acids Res 31:3320–3323
Fitch WM (1971) Syst Zool 20:406–416
Galtier N, Gouy M, Gautier C (1996) Comput Appl Biosci 12:543–548
Saitou N, Nei M (1987) Mol Biol Evol 4:406–425
Felsenstein J (1985) Evolution 39:783–791
Guex N, Peitsch MC (1997) Electrophoresis 18:2714–272321
Littlechild JA, Garcia-Rodriguez E, Dalby AR, Isupov MN (2002) J Mol Recognit 15:291–296
Bhattacharya D, Medlin L (1998) Plant Physiol 116:9–15
Baldauf SL, Roger AJ, Wenk-Siefert I, Doolittle WF (2000) Science 290:972–977
Draisma SGA, Peters AF, Fletcher RL (2003) In: Norton TA (ed) Out of the past. The British phycological society, pp87–102
Almeida M, Filipe S, Humanes M, Maia MF, Melo R, Severino N, da Silva JAL, Frausto da Silva JJR, Wever R (2001) Phytochemistry 57:633–642
Verschueren KH, Kingma J, Rozeboom HJ, Kalk KH, Janssen DB, Dijkstra BW (1993) Biochemistry 32:9031–9037
Machius M, Wiegand G, Huber R (1995) J Mol Biol 246:545–559
Fukuyama K, Sato K, Itakura H, Takahashi S, Hosoya T (1997) J Biol Chem 272:5752–5756
Fiedler TJ, Davey CA, Fenna RE (2000) J Biol Chem 275:11964–11971
Saenko GN, Kravtsova YY, Ivanenko VV, Sheludko S I (1978) Mar Biol 47:243–250
Acknowledgements
This work was supported in part by the European Commission “GROWTH Program”, Research Project AB, Algal Bioadhesives, G5RD-CT-2001-00542, by a Ph.D. grant from the Région Bretagne (to C.C.) and a Ph.D. grant from Bruker–Daltonic (to E.W.). We are grateful to Agilent Technologies for providing the capillary LC system for MS analyses and to Marc Cock for critical reading of the manuscript.
Author information
Authors and Affiliations
Corresponding author
Additional information
The nucleotide sequence reported in this paper has been submitted to the EBI Data Bank with accession no. AJ619804.
Rights and permissions
About this article
Cite this article
Colin, C., Leblanc, C., Michel, G. et al. Vanadium-dependent iodoperoxidases in Laminaria digitata, a novel biochemical function diverging from brown algal bromoperoxidases. J Biol Inorg Chem 10, 156–166 (2005). https://doi.org/10.1007/s00775-005-0626-8
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00775-005-0626-8