Thioredoxin-related protein of 14 kDa is an efficient L-cystine reductase and S-denitrosylase.
Thioredoxin-related protein of 14 kDa (TRP14, also called TXNDC17 for thioredoxin domain containing 17, or TXNL5 for thioredoxin-like 5) is an evolutionarily well-conserved member of the thioredoxin (Trx)-fold protein family that lacks activity with classical Trx1 substrates. However, we discovered here that human TRP14 has a high enzymatic activity in reduction of l-cystine, where the catalytic efficiency (2,217 min(-1)⋅µM(-1)) coupled to Trx reductase 1 (TrxR1) using NADPH was fivefold higher compared with Trx1 (418 min(-1)⋅µM(-1)). Moreover, the l-cystine reduction with TRP14 was in contrast to that of Trx1 fully maintained in the presence of a protein disulfide substrate of Trx1 such as insulin, suggesting that TRP14 is a more dedicated l-cystine reductase compared with Trx1. We also found that TRP14 is an efficient S-denitrosylase with similar efficiency as Trx1 in catalyzing TrxR1-dependent denitrosylation of S-nitrosylated glutathione or of HEK293 cell-derived S-nitrosoproteins. Consequently, nitrosylated and thereby inactivated caspase 3 or cathepsin B could be reactivated through either Trx1- or TRP14-catalyzed denitrosylation reactions. TRP14 was also, in contrast to Trx1, completely resistant to inactivation by high concentrations of hydrogen peroxide. The oxidoreductase activities of TRP14 thereby complement those of Trx1 and must therefore be considered for the full understanding of enzymatic control of cellular thiols and nitrosothiols.
Division of Biochemistry, Department of Medical Biochemistry and Biophysics and.
Irina
I
Pader
Division of Biochemistry, Department of Medical Biochemistry and Biophysics and.
Rajib
R
Sengupta
Division of Biochemistry, Department of Medical Biochemistry and Biophysics and.
Marcus
M
Cebula
Division of Biochemistry, Department of Medical Biochemistry and Biophysics and.
Jianqiang
J
Xu
Department of Physiology and Pharmacology, Karolinska Institutet, SE-171 77 Stockholm, Sweden.
Jon O
JO
Lundberg
Division of Biochemistry, Department of Medical Biochemistry and Biophysics and.
Arne
A
Holmgren
Division of Biochemistry, Department of Medical Biochemistry and Biophysics and.
Katarina
K
Johansson
Division of Biochemistry, Department of Medical Biochemistry and Biophysics and elias.arner@ki.se.
Elias S J
ES
Arnér
D002294:Carcinoma, Squamous Cell; D003545:Cysteine / Q000378:metabolism; D003553:Cystine / Q000378:metabolism*; D004789:Enzyme Activation / Q000502:physiology; D005978:Glutathione / Q000378:metabolism; D057809:HEK293 Cells; D019073:HT29 Cells; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D008175:Lung Neoplasms; D009247:NADH, NADPH Oxidoreductases / Q000235:genetics / Q000378:metabolism; D009249:NADP / Q000378:metabolism; D009569:Nitric Oxide / Q000378:metabolism; D016877:Oxidants / Q000494:pharmacology; D010084:Oxidation-Reduction; D018384:Oxidative Stress / Q000502:physiology*; D013379:Substrate Specificity; D013455:Sulfur / Q000378:metabolism; D054481:Thioredoxin Reductase 1 / Q000235:genetics / Q000378:metabolism*; D013879:Thioredoxins / Q000235:genetics / Q000378:metabolism*
D016877:Oxidants; C482341:TXNDC17 protein, human; D009569:Nitric Oxide; D003553:Cystine; D013879:Thioredoxins; D009249:NADP; D013455:Sulfur; D006861:Hydrogen Peroxide; D009247:NADH, NADPH Oxidoreductases; C019976:Cystine reductase; C516124:TXNRD1 protein, human; D054481:Thioredoxin Reductase 1; D005978:Glutathione; D003545:Cysteine
nitric oxide; oxidative stress; redox regulation; sulfur metabolism