Pholasin: a new chemiluminescent probe for the detection of chloramines derived from human phagocytes.

Abstract

We have previously reported that normal human polymorphonuclear neutrophils (PMN) release taurine-chloramine, a long-lived oxidant, in response to stimulation by phorbol myristate acetate (PMA) or opsonized zymosan in the presence of exogenous taurine. We now describe a new, simple, and highly sensitive method for the detection of chloramines, including taurine-chloramine, using the chemiluminescent probe Pholasin, the luciferin of the mollusc Pholas dactylus. Taurine-chloramine (N-chlorotaurine) detection was assessed with both a colorimetric method (based on the oxidation of potassium iodide) and with the Pholasin-dependent chemiluminescence (CL) method. The taurine-chloramine concentration in PMN supernatants determined using the potassium iodide (KI) method correlated closely with Pholasin-dependent CL. This CL was also assessed in nonoxidative conditions. No taurine-chloramine was detected in supernatants of lymphocytes and PMN from patients with an oxidative burst defect (chronic granulomatous disease, CGD) with the KI method, but Pholasin-dependent CL was consistently observed. The use of methionine, a specific chloramine scavenger in our incubation conditions, allowed us to define a methionine-inhibitable fraction of Pholasin-dependent CL (i.e., chloramine-induced CL).

• Authors: V Witko-Sarsat, A T Nguyen, J Knight, B Descamps-Latscha
• Journal: Free radical biology & medicine (Free Radic Biol Med) Vol. 13 Issue 1 Pg. 83-8 ( 1992) ISSN: 0891-5849 [Print] United States
• PMID: 1628856 (Publication Type: Comparative Study, Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Chloramines
• Free Radicals
• Indicators and Reagents
• Taurine
• N-chlorotaurine
• Firefly Luciferin
• pholasin

Topics

• Child
• Chloramines (metabolism)
• Firefly Luciferin
• Free Radicals
• Granulomatous Disease, Chronic (metabolism)
• Humans
• In Vitro Techniques
• Indicators and Reagents
• Luminescent Measurements
• Neutrophils (metabolism)
• Phagocytes (metabolism)
• Taurine (analogs & derivatives, metabolism)