Effects of heme proteins on nitric oxide levels and cell viability in isolated PMNs: a mechanism of toxicity

Cassidy, Richard A.; Burleson, David G.; Delgado, Ángel Acuña; Köhler, D; Salin, Marvin L.; Pruitt, Basil A.

doi:10.1002/jlb.67.3.357

Cited by 9 publications

(18 citation statements)

References 63 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…It remains to be established whether the recovery of bovine catalase from the cells reflects an actual internalization. Other heme proteins have been found to be internalized into neutrophils (oxyhemoglobin) [44] or transcytosed into the intact endothelium (MPO) [45].…”

Section: Discussionmentioning

confidence: 99%

Identification of proteins in activated human neutrophils susceptible to tyrosyl radical attack. A proteomic study using a tyrosylating fluorophore

et al. 2004

View full text Add to dashboard Cite

Tyrosyl radicals cross-linked to protein tyrosine residues (tyrosylated proteins) represent hallmarks of neutrophil-mediated injury at the inflammatory locus. Yet the proteins targeted by tyrosyl radicals in an intact cellular system remain to be elucidated. Here, we show that tyrosyl radicals generated by human neutrophils after activation by phorbol 12-myristate 13-acetate (PMA), interferon-g (IFN-g) or TNF-a could act in an autocrine manner by cross-linking to endogenous proteins. We have identified the tyrosylated proteins by using a membraneimpermeable tyrosine analogue, tyramine coupled to fluorescein (TyrFluo), in combination with proteomics techniques. Confocal microscopy images indicated that initially the tyrosylated proteins were localized in patches at the cell surface to become internalized subsequently. In the neutrophil membrane-associated proteome, lactoferrin was the prime target of tyrosylation. Out of three isoforms identified, an 80 kDa neutral isoform was tyrosylated more extensively than the 85 kD basic isoform, particularly after PMA activation. Although all three stimuli induced tyrosylation of the filamentous component vimentin, additional tyrosylated vimentin fragments were detected after IFN-g-and TNF-a-stimulation. Moreover, upon activation the bulk of vimentin behaved as a dimer (M r 120 kDa) being slightly tyrosylated, yet phosphorylated at Thr-425 possibly as a requirement for its externalization. Unexpectedly, bovine catalase added to end tyrosyl radicals formation was detected as a highly tyrosylated neutrophil-associated protein. A moderate stimulus-dependent tyrosylation of ATP synthaseb, a-enolase, glyceraldehyde 3-phosphate dehydrogenase, cytokeratin-10, filamin-A, and annexin-I was also observed. When the membrane-permeable probe (acetylTyrFluo) was used, protein tyrosylation was not observed indicating that the intracellular proteins were well protected against oxidative attack. This study shows that human neutrophils can modulate their proteome via a tyrosine oxidation pathway induced by pro-inflammatory mediators.

show abstract

Section: Discussionmentioning

confidence: 99%

Identification of proteins in activated human neutrophils susceptible to tyrosyl radical attack. A proteomic study using a tyrosylating fluorophore

et al. 2004

View full text Add to dashboard Cite

show abstract

“…Polymorphonuclear leukocytes (PMNs) and lymphocytes were obtained from EDTA-preserved venous blood by layering blood over Polymorphprep (Nycomed Pharma, Oslo, Norway) followed by centrifugation as previously described [30]. The purity of PMNs and lymphocytes was greater than 88 and 95%, respectively, and their viability, as determined by trypan-blue exclusion, was greater than 90%.…”

Section: In-vitro Studiesmentioning

confidence: 99%

“…The effect of oxidants, derived from HE-treated PMNs, on the membrane integrity of neighboring lymphocyte target cells was determined by propidium iodide (PI) exclusion in a mixed PMN/lymphocyte preparation using 100,000 lymphocytes and PMN/lymphocyte ratios of 0/1, 2/1, 4/1, and 10/1) [30]. To obtain these ratios, varying numbers of PMNs (prepared at a density of 300,000/100 ll HBSS) from four different treatment preparations were mixed with lymphocytes (100,000/ 100 ll HBSS) from the same donor and incubated for 40 min at 37°C.…”

Section: Effect Of He-treated Pmns On Cell-membrane Integrity Of Co-imentioning

confidence: 99%

“…Effect of HE on DNA strand breaks HE-induced DNA strand breaks in PMNs were measured by flow cytometry using the terminal deoxynucleotidyl transferase-mediated nick end-labeling assay (TUNEL) [30] [In-Situ Cell Death Detection Kit (Boehringer Mannheim, Indianapolis, Indiana) according to manufacturer's instructions]. PMNs were incubated at 37°C with and without the previously-mentioned concentrations of L L -NMMA, 17b-E 2 receptor antagonists, or TNFa, and varying concentrations of HE as described above for the HE-induced DCFH oxidation incubations, with the exception that HE incubation time was 2 h, instead of 40 min.…”

Section: Effect Of He (Or 17b-e 2 ) On Intracellular Oxidantsmentioning

confidence: 99%

“…Intracellular levels of oxidants in PMNs were measured using a previously reported flow cytometric procedure [30]. Briefly, PMNs were loaded with 2,7-dichlorodihydrofluorescein diacetate (DCFH-DA)(Kodak, Rochester, New York) at 37°C for 15 min in a gently rocking water bath followed by a 10 min incubation without and with the specific NO synthase inhibitor N-methyl-L Larginine (L L -NMMA) ( and with inhibitors or TNFa were subsequently incubated with varying concentrations of HE (zero and 0.01-10 nM) (Ultra Scientific, Kingstown, Rhode Island) at 37°C for 40 min.…”

Section: Effect Of He (Or 17b-e 2 ) On Intracellular Oxidantsmentioning

confidence: 99%

See 2 more Smart Citations

The link between the insecticide heptachlor epoxide, estradiol, and breast cancer

Cassidy¹,

Sridhar

Vaughan

2005

Breast Cancer Res Treat

Self Cite

View full text Add to dashboard Cite

Given the suspected effects of estrogens on breast cancer, xenoestrogenic insecticides may be a risk factor. Studies of the weak xenoestrogen, 1,1-dichloro-2,2-bis(p-chlorophenyl)ethylene (DDE), have failed to demonstrate a causal relationship, though another estrogenic organochlorine insecticide, dieldrin, belonging to the cyclodiene family, has recently been linked to breast cancer. Other cyclodienes such as heptachlor epoxide (HE) and oxychlordane (OC) present in breast tissue have not been evaluated as rigorously, presumably due to their lower concentration and lower recovery using solvent extraction procedures. We used sparging extraction coupled with gas chromatography to determine the levels of HE, OC, and DDE in adipose tissue within breast biopsies in a series of 34 women evaluated for breast abnormality. Of the three insecticides tested, only HE (p=0.007) was positively associated with prevalence of breast cancer in the biopsies. In rapid, non-genomic studies using isolated human leukocytes, flow cytometric methods were used to measure HE-induced oxidants and DNA damage. These studies indicated that HE, at concentrations similar to those in breast biopsies, induced an inverted-U increase in intracellular oxidants and DNA strand breaks [both blocked by specific nitric oxide- (NO-) synthesis blockade withL: -NMMA] in human polymorphonuclear leukocytes (PMNs). HE-treated PMNs also induced damage to surrounding lymphocytes in mixed-leukocyte incubations (also inhibited by NO blockade). The HE-induced changes in NO were inhibited by 17beta-estradiol-(17beta-E2) receptor antagonists and were mimicked by similar concentrations of 17beta-E2. The addition of tumor necrosis factor-alpha (TNF-alpha) increased intracellular oxidants and DNA damage and shifted the responses to lower HE concentrations. This study, along with others, suggests that HE-induced NO production may contribute to initiation, promotion, and progression of cancer.

show abstract

Hemoglobin S Polymerization and Red Cell Membrane Changes

Kuypers¹

2014

Hematology/Oncology Clinics of North America

View full text Add to dashboard Cite

Effects of heme proteins on nitric oxide levels and cell viability in isolated PMNs: a mechanism of toxicity

Cited by 9 publications

References 63 publications

Identification of proteins in activated human neutrophils susceptible to tyrosyl radical attack. A proteomic study using a tyrosylating fluorophore

Identification of proteins in activated human neutrophils susceptible to tyrosyl radical attack. A proteomic study using a tyrosylating fluorophore

The link between the insecticide heptachlor epoxide, estradiol, and breast cancer

Hemoglobin S Polymerization and Red Cell Membrane Changes

Contact Info

Product

Resources

About