UNCEIN MAY BE A POTENTIAL TARGET FOR SULFUR MUSTARD ALKYLATION

Abstract

Severe blister formation in theepidermal and dermal junction is one of the characteristic cutaneous lesions caused by bis-2-chloroethyl sulfide (sulfur mustard, HD), a bifunctional alkylating agent. Uncein is a newly discovered anchoring filament-associated antigen that has been shown to be undetectable in all forms of junctional epidermolysis bullosa, therefore suggesting its role in maintaining the integrity of the epidermal-dermal basement membrane zone. To test uncein as a potential target for HD alkylation in HD-induced vesication, uncein was biosynthetically labeled with 35 S-methionine and purified by immunoprecipitation with a 19-DEJ-1 monoclonal antibody from the medium of normal human keratinocyte (NHEK) cultures. The 3 5 S-labeled uncein was incubated with 50 muL of 10.0 mg/mL of HD in ethanol (ETOH) or ETOH, which served as the control in 50 mM Tris-Cl buffer (pH 7.4) for 2 h. In addition, 10.0 mg/mL of HD was incubated with uncein pretreated with 3 mM sodium thiosulfate, an HD scavenger. Unce...