Purification, properties and primary structure of thioredoxin from Aspergillus nidulans.
This paper reports the purification and the properties of a thioredoxin from the fungus Aspergillus nidulans. This thioredoxin is an acidic protein which exhibits an unusual fluorescence emission spectrum, characterized by a high contribution of tyrosine residues. Thioredoxin from A. nidulans cannot serve as a substrate for Escherichia coli thioredoxin reductase. Corn NADP-malate dehydrogenase is activated by this thioredoxin in the presence of dithiothreitol, while fructose-1,6-bisphosphatase is not. The amino acid sequence of Aspergillus thioredoxin was determined by automated Edman degradation after cleavage with trypsin, SV8 protease, chymotrypsin and cyanogen bromide. The masses of tryptic peptides were verified by plasma-desorption mass spectrometry. The mass of the protein was determined by electrospray mass spectrometry and shown to be in agreement with the calculated mass derived from the sequence (M(r) = 11,564). Compared to thioredoxins from other sources, the protein from A. nidulans displays a maximal sequence similarity with that from yeast (45%).
Le Marechal
P
P
Laboratoire de Physiologie Végétale Moléculaire, Université Paris-Sud, Orsay, France.
Hoang
B M
BM
Schmitter
J M
JM
Van Dorsselaer
A
A
Decottignies
P
P
D000595:Amino Acid Sequence; D001233:Aspergillus nidulans / Q000737:chemistry*; D004229:Dithiothreitol / Q000494:pharmacology; D004926:Escherichia coli / Q000201:enzymology; D006863:Hydrogen-Ion Concentration; D007526:Isoelectric Point; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009249:NADP / Q000494:pharmacology; D010446:Peptide Fragments / Q000737:chemistry; D017386:Sequence Homology, Amino Acid; D013050:Spectrometry, Fluorescence; D013880:Thioredoxin-Disulfide Reductase / Q000378:metabolism; D013879:Thioredoxins / Q000737:chemistry* / Q000302:isolation & purification
D010446:Peptide Fragments; D013879:Thioredoxins; D009249:NADP; D013880:Thioredoxin-Disulfide Reductase; D004229:Dithiothreitol