Myeloperoxidase interacts with endothelial cell-surface cytokeratin 1 and modulates bradykinin production by the plasma Kallikrein-Kinin system.
During an inflammatory state, functional myeloperoxidase (MPO) is released into the vessel as a result of intravascular neutrophil degradation. One mechanism of resulting cellular injury involves endothelial internalization of MPO, which causes oxidative damage and impairs endothelial signaling. We report the discovery of a protein that facilitates MPO internalization, cytokeratin 1 (CK1), identified using affinity chromatography and mass spectrometry. CK1 interacts with MPO in vitro, even in the presence of 100% human plasma, thus substantiating biological relevance. Immunofluorescent microscopy confirmed that MPO added to endothelial cells can co-localize with endogenously expressed CK1. CK1 acts as a scaffolding protein for the assembly of the vasoregulatory plasma kallikrein-kinin system; thus we explored whether MPO and high molecular weight kininogen (HK) reside on CK1 together or whether they compete for binding. The data support cooperative binding of MPO and HK on cells such that MPO masked the plasma kallikrein cleavage site on HK, and MPO-generated oxidants caused inactivation of both HK and kallikrein. Collectively, interactions between MPO and the components of the plasma kallikrein-kinin system resulted in decreased bradykinin production. This study identifies CK1 as a facilitator of MPO-mediated vascular responses and thus provides a new paradigm by which MPO affects vasoregulatory systems.
Astern
Joshua M
JM
Burnett-Womack, UNC Kidney Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599-7155, USA. jastern@med.unc.edu
Pendergraft
William F
WF
Falk
Ronald J
RJ
Jennette
J Charles
JC
Schmaier
Alvin H
AH
Mahdi
Fakhri
F
Preston
Gloria A
GA
D001920:Bradykinin / Q000096:biosynthesis*; D002352:Carrier Proteins; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000378:metabolism* / Q000648:ultrastructure; D004730:Endothelium, Vascular / Q000378:metabolism*; D006801:Humans; D016234:Kallikrein-Kinin System / Q000502:physiology*; D053556:Keratin-1 / Q000378:metabolism / Q000502:physiology*; D053534:Keratin-9 / Q000378:metabolism; D019679:Kininogen, High-Molecular-Weight / Q000378:metabolism; D046911:Macromolecular Substances / Q000378:metabolism; D009195:Peroxidase / Q000502:physiology*
D002352:Carrier Proteins; D053556:Keratin-1; D053534:Keratin-9; D019679:Kininogen, High-Molecular-Weight; D046911:Macromolecular Substances; D009195:Peroxidase; D001920:Bradykinin