Secondary structure and protein folding of recombinant chloroplastic thioredoxin Ch2 from the green alga Chlamydomonas reinhardtii as determined by 1H NMR.
The recombinant form of the chloroplastic thioredoxin Ch2 from the green alga Chlamydomonas reinhardtii [Jacquot et al. (1992) Nucleic Acids Res. 20, 617] that preferentially activates the NADP dependent malate dehydrogenase [EC 1.1.1.82] (m-type thioredoxin) through a light promoted reductive system, has been subjected to an extensive two-dimensional 1H NMR analysis. A complete 1H NMR assignment of the resonance lines in both the oxidized and the reduced states at pH 5.8 has been obtained allowing the recognition of the secondary structure patterns and the global protein folding. The single polypeptide chain, made of 106 residues plus one additional Met located at the N-terminal position (11.6 kDa) due to the protein expression system, folds into a pattern characteristic of the open twisted alpha/beta structures already found for Escherichia coli and human thioredoxins for which the protein shares 46 and 20% of sequence identity, respectively. The open alpha/beta structure is made of 5 beta-sheets associated in a parallel (beta 1 to beta 3) and anti parallel manner (beta 3 to beta 5) and surrounded by 4 helices. This represents the first structural exploratory study of the ubiquitous oxido-reductase thioredoxins in a photosynthetic living system.
Lancelin
J M
JM
Institut de Biologie Structurale CEA-CNRS, Grenoble, France.
Stein
M
M
Jacquot
J P
JP
D000596:Amino Acids / Q000737:chemistry; D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry; D016825:Chlamydomonas reinhardtii; D002736:Chloroplasts / Q000737:chemistry*; D004926:Escherichia coli / Q000737:chemistry; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D010084:Oxidation-Reduction; D010940:Plant Proteins / Q000737:chemistry*; D017510:Protein Folding; D017433:Protein Structure, Secondary; D011522:Protons; D011994:Recombinant Proteins / Q000737:chemistry; D017386:Sequence Homology, Amino Acid; D013879:Thioredoxins / Q000737:chemistry*
D000596:Amino Acids; D001426:Bacterial Proteins; D010940:Plant Proteins; D011522:Protons; D011994:Recombinant Proteins; C073081:thioredoxin Ch2 protein, Chlamydomonas reinhardtii; D013879:Thioredoxins