Myeloperoxidase inactivates TIMP-1 by oxidizing its N-terminal cysteine residue: an oxidative mechanism for regulating proteolysis during inflammation.
An imbalance between the proteolytic activity of matrix metalloproteinases (MMPs) and the activity of tissue inhibitors of metalloproteinases (TIMPs) is implicated in tissue injury during inflammation. The N-terminal cysteine of TIMP-1 plays a key role in the inhibitory activity of the protein because it coordinates the essential catalytic Zn2+ of the MMP, preventing the metal ion from functioning. An important mechanism for controlling the interaction of TIMPs with MMPs might involve hypochlorous acid (HOCl), a potent oxidant produced by the myeloperoxidase (MPO) system of phagocytes. Here, we show that HOCl generated by the MPO-H2O2-chloride system inactivates TIMP-1 by oxidizing its N-terminal cysteine. The product is a novel 2-oxo acid. Liquid chromatography-mass spectrometry and tandem mass spectrometry analyses demonstrated that methionine and N-terminal cysteine residues were rapidly oxidized by MPO-derived HOCl but only oxidation of the N-terminal cysteine of TIMP-1 correlated well with loss of inhibitory activity. Importantly, we detected the signature 2-oxo-acid N-terminal peptide in tryptic digests of bronchoalveolar lavage fluid from patients with acute respiratory distress syndrome, demonstrating that TIMP-1 oxidation occurs in vivo. Loss of the N-terminal amino group and disulfide structure are crucial for preventing TIMP-1 from inhibiting MMPs. Our findings suggest that pericellular production of HOCl by phagocytes is a pathogenic mechanism for impairing TIMP-1 activity during inflammation.
Wang
Yi
Y
Department of Medicine, University of Washington, Fred Hutchinson Cancer Research Center, Seattle, Washington 98195, USA.
Rosen
Henry
H
Madtes
David K
DK
Shao
Baohai
B
Martin
Thomas R
TR
Heinecke
Jay W
JW
Fu
Xiaoyun
X
D000595:Amino Acid Sequence; D006801:Humans; D006997:Hypochlorous Acid / Q000378:metabolism; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D009195:Peroxidase / Q000378:metabolism*; D010586:Phagocytes / Q000378:metabolism; D012128:Respiratory Distress Syndrome / Q000378:metabolism*; D053719:Tandem Mass Spectrometry; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000737:chemistry* / Q000378:metabolism*
D019715:Tissue Inhibitor of Metalloproteinase-1; D006997:Hypochlorous Acid; D009195:Peroxidase