Nitrogen monoxide (NO) storage and transport by dinitrosyl-dithiol-iron complexes: long-lived NO that is trafficked by interacting proteins.
ABSTRACT Nitrogen monoxide (NO) markedly affects intracellular iron metabolism, and recent studies have shown that molecules traditionally involved in drug resistance, namely GST and MRP1 (multidrug resistance-associated protein 1), are critical molecular players in this process. This is mediated by interaction of these proteins with dinitrosyl-dithiol-iron complexes (Watts, R. N., Hawkins, C., Ponka, P., and Richardson, D. R. (2006) Proc. Natl. Acad. Sci. U.S.A. 103, 7670-7675; Lok, H. C., Suryo Rahmanto, Y., Hawkins, C. L., Kalinowski, D. S., Morrow, C. S., Townsend, A. J., Ponka, P., and Richardson, D. R. (2012) J. Biol. Chem. 287, 607-618). These complexes are bioavailable, have a markedly longer half-life compared with free NO, and form in cells after an interaction between iron, NO, and glutathione. The generation of dinitrosyl-dithiol-iron complexes acts as a common currency for NO transport and storage by MRP1 and GST P1-1, respectively. Understanding the biological trafficking mechanisms involved in the metabolism of NO is vital for elucidating its many roles in cellular signaling and cytotoxicity and for development of new therapeutic targets.
Article: Glutathione transferase P1 interacts strongly with the inner leaflet of the plasma membrane.[show abstract] [hide abstract]
ABSTRACT: GSH transferases (GSTs) are a superfamily of proteins best known for detoxifying harmful electrophilic compounds by catalyzing their conjugation with GSH. GSTP1 is the most prevalent and widely distributed GST in human tissues, helping to detoxify a diverse array of carcinogens and drugs. In contrast with its protective role, overexpression of GSTP1 in a variety of malignancies is associated with a poor prognosis due to failure of chemotherapy. Although GSTP1 is classified as a cytosolic GST, we discovered previously that it is associated with the plasma membrane of the small cell lung cancer cell lines, H69 and H69AR. In the current study, endogenous and overexpressed GSTP1 in human embryonic kidney (HEK) 293 and MCF-7 cell lines, respectively, were found also to associate with the plasma membrane, indicating that this interaction is not unique to H69 and H69AR cells. GSTP1 immunostaining in HEK293 and MCF7-GSTP1 cells only occurred under permeabilized conditions, suggesting that GSTP1 is associated with the intracellular surface of the plasma membrane. Cell surface biotinylation studies confirmed this finding. Immunogold electron microscopy revealed the presence of GSTP1 in close proximity to the plasma membrane. GSTP1 was not dissociated from plasma membrane sheets by high salt [potassium iodide (KI; 1 M) or KI/EDTA (1 M/2 mM)] or alkaline Na(2)CO(3) (100 mM, pH 11.4), conditions known to strip peripherally associated membrane proteins. Thus, we report for the first time that GSTP1 is associated with the inner leaflet of the plasma membrane through a remarkably strong interaction.Drug metabolism and disposition: the biological fate of chemicals 04/2011; 39(7):1122-6. · 3.74 Impact Factor
Article: Pancreatic endocrine responses to substance P and calcitonin gene-related peptide in conscious calves.[show abstract] [hide abstract]
ABSTRACT: Both substance P (SP) and calcitonin gene-related peptide (CGRP; 0.13 micrograms.min-1.kg-1 for 10 min) produced a significant rise in arterial plasma pancreatic polypeptide (PP) concentration, and additive responses were obtained when both peptides were infused simultaneously and/or with acetylcholine (0.7 micrograms.min-1.kg-1 ia). The PP response to SP was abolished by intravenous infusions of glucose, whereas those to CGRP and acetylcholine were not significantly affected. Neither SP nor CGRP had any effect on plasma insulin concentration, either in the presence or absence of exogenous glucose, whether infused singly or together, or in the presence of acetylcholine. SP, but not CGRP, produced a small but statistically significant rise in mean plasma glucagon concentration when infused together with acetylcholine. These results suggest that SP and CGRP may modulate the secretion of PP and glucagon in the normal conscious calf but not that of insulin. It is also possible that SP modulates secretion of pancreatic glucagon in these animals.The American journal of physiology 01/1995; 267(6 Pt 1):E847-52.
Article: The potent vasodilating and guanylyl cyclase activating dinitrosyl-iron(II) complex is stored in a protein-bound form in vascular tissue and is released by thiols.[show abstract] [hide abstract]
ABSTRACT: We studied the biological activity, stability and interaction of dinitrosyl-iron(II)-L-cysteine with vascular tissue. Dinitrosyl-iron(II)-L-cysteine was a potent activator of purified soluble guanylyl cyclase (EC50 10 nM with and 100 nM without superoxide dismutase) and relaxed noradrenaline-precontracted segments of endothelium-denuded rabbit femoral artery (EC50 10 nM superoxide dismutase). Pre-incubation (5 min; 310 K) of endothelium-denuded rabbit aortic segments with dinitrosyl-iron(II)-L-cysteine (0.036-3.6 mM) resulted in a concentration-dependent formation of a dinitrosyl-iron(II) complex with protein thiol groups, as detected by ESR spectroscopy. While the complex with proteins was stable for 2 h at 310 K, dinitrosyl-iron(II)-L-cysteine in aqueous solution (36-360 microM) decomposed completely within 15 min, as indicated by disappearance of its isotropic ESR signal at gav = 2.03 (293 K). Aortic segments pre-incubated with dinitrosyl-iron(II)-L-cysteine released a labile vasodilating and guanylyl cyclase activating factor. Perfusion of these segments with N-acetyl-L-cysteine resulted in the generation of a low molecular weight dinitrosyl-iron(II)-dithiolate from the dinitrosyl-iron(II) complex with proteins, as revealed by the shape change of the ESR signal at 293 K. The low molecular weight dinitrosyl-iron(II)-dithiolate accounted for an enhanced guanylyl cyclase activation and vasodilation induced by the aortic effluent. We conclude that nitric oxide (NO) produced by, or acting on vascular cells can be stabilized and stored as a dinitrosyl-iron(II) complex with protein thiols, and can be released from cells in the form of a low molecular weight dinitrosyl-iron(II)-dithiolate by intra- and extracellular thiols.FEBS Letters 01/1992; 294(3):252-6. · 3.54 Impact Factor