caa a22 4500 445322101 CHVBK 20180317142707.0 cr unu---uuuuu 170323e20110401xx s 000 0 eng 10.1007/s10534-010-9394-1 doi (NATIONALLICENCE)springer-10.1007/s10534-010-9394-1 Ferrous iron is found in mesenteric lymph bound to TIMP-2 following hemorrhage/resuscitation [Elektronische Daten] [James Atkins, Nikolai Gorbunov, Valerie Trabosh, Rachel Van Duyne, Fatah Kashanchi, Andrei Komarov] Extracellular iron has been implicated in the pathogenesis of post-injury organ failure. However, the source(s) and biochemical species of this iron have not been identified. Based upon evidence that distant organ injury results from an increase in intestinal permeability, we looked for ferrous iron in mesenteric lymph in anesthetized rats undergoing hemorrhage and fluid resuscitation (H/R). Ferrous iron increased in lymph from 4.7nmol/mg of protein prior to hemorrhage to 86.6nmol/mg during resuscitation. Utilizing immuno-spin trapping in protein fractions that were rich in iron, we tentatively indentified protein carrier(s) of ferrous iron by MALDI-TOF MS. One of the identified proteins was the metalloproteinase (MMP) inhibitor, TIMP-2. Antibody to TIMP-2 immunoprecipitated 74% of the ferrozine detectable iron in its protein fraction. TIMP-2 binds iron in vitro at pH 6.3, which is typical of conditions in the mesentery during hemorrhage, but it retains the ability to inhibit the metalloproteases MMP-2 and MMP-9. In summary, there is a large increase in extracellular ferrous iron in the gut in H/R demonstrating dysregulation of iron homeostasis. We have identified, for the first time, the binding of extracellular iron to TIMP-2. Springer Science+Business Media, LLC (outside the USA), 2011 Ferrous iron nationallicence TIMP-2 nationallicence Hemorrhage nationallicence Extracellular nationallicence A&G bead : Protein A/G Sepharose beads nationallicence ALI : Acute lung injury nationallicence Brij-35 : Ethoxylated dodecyl alcohol nationallicence Dcytb : Duodenal cytochrome B nationallicence DMPO : 5,5-Dimethyl-1-pyrroline N-oxide nationallicence DTNB : 5,5′-Dithiobis-(2-nitrobenzoic acid) nationallicence DTT : Dithiothreitol nationallicence DUSP-12 : Dual specificity protein phosphatase 12 nationallicence EDTA : Ethylenediaminetetraacetic acid nationallicence ELISA : Enzyme-linked immunosorbent assay nationallicence EPR : Electron paramagnetic resonance nationallicence HEPES : 4-(2-Hydroxyethyl)-1-piperazineethanesulfonic acid nationallicence H/R : Hemorrhage/resuscitation nationallicence IP : Immunoprecipitation nationallicence MALDI-TOF MS : Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry nationallicence MI/R : Mesenteric ischemia/reperfusion nationallicence MMP : Matrix metalloproteinase nationallicence NP-40 : Nonyl phenoxypolyethoxylethanol nationallicence PBS : Phosphate buffered saline nationallicence PMSF : Phenylmethanesulfonylfluoride nationallicence SDS-PAGE : Sodium dodecyl sulfate polyacrylamide gel electrophoresis nationallicence TIMP-2 : Tissue inhibitor of metalloproteinases-2 nationallicence WLE : Whole lymph extract nationallicence Atkins James Closed Head Brain Injury Branch, Center for Military Psychiatry and Neuroscience, Walter Reed Army Institute of Research, 503 Robert Grant Ave., 20910-7500, Silver Spring, MD, USA aut Gorbunov Nikolai The Henry M. Jackson Foundation for the Advancement of Military Medicine, Inc., 20889, Bethesda, MD, USA aut Trabosh Valerie Closed Head Brain Injury Branch, Center for Military Psychiatry and Neuroscience, Walter Reed Army Institute of Research, 503 Robert Grant Ave., 20910-7500, Silver Spring, MD, USA aut Van Duyne Rachel Department of Molecular and Microbiology, National Center for Biodefense and Infectious Diseases, George Mason University, 20110, Manassas, VA, USA aut Kashanchi Fatah Department of Molecular and Microbiology, National Center for Biodefense and Infectious Diseases, George Mason University, 20110, Manassas, VA, USA aut Komarov Andrei The George Washington University Medical Center, 20037, Washington, DC, USA aut BioMetals Springer Netherlands 24/2(2011-04-01), 279-289 0966-0844 24:2<279 2011 24 10534 https://doi.org/10.1007/s10534-010-9394-1 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10534-010-9394-1 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Atkins James Closed Head Brain Injury Branch, Center for Military Psychiatry and Neuroscience, Walter Reed Army Institute of Research, 503 Robert Grant Ave., 20910-7500, Silver Spring, MD, USA aut NATIONALLICENCE 700 1- Gorbunov Nikolai The Henry M. Jackson Foundation for the Advancement of Military Medicine, Inc., 20889, Bethesda, MD, USA aut NATIONALLICENCE 700 1- Trabosh Valerie Closed Head Brain Injury Branch, Center for Military Psychiatry and Neuroscience, Walter Reed Army Institute of Research, 503 Robert Grant Ave., 20910-7500, Silver Spring, MD, USA aut NATIONALLICENCE 700 1- Van Duyne Rachel Department of Molecular and Microbiology, National Center for Biodefense and Infectious Diseases, George Mason University, 20110, Manassas, VA, USA aut NATIONALLICENCE 700 1- Kashanchi Fatah Department of Molecular and Microbiology, National Center for Biodefense and Infectious Diseases, George Mason University, 20110, Manassas, VA, USA aut NATIONALLICENCE 700 1- Komarov Andrei The George Washington University Medical Center, 20037, Washington, DC, USA aut NATIONALLICENCE 773 0- BioMetals Springer Netherlands 24/2(2011-04-01), 279-289 0966-0844 24:2<279 2011 24 10534 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer