Characterization of elastase-like enzymes in various blistering diseases

Blister fluid samples were collected from suction induced control blisters and from spontaneous blisters from various blistering diseases for the measurement of elastase-like enzyme activities using synthetic succinyl-L-(alanyl)3-paranitroanilide (SAPNA) and alanyl paranitroanilide (ala-PNA) as the substrates. The blister fluids derived from bullous pemphigoid, pyoderma or epidermolysis bullosa dystroficans lesions contained higher levels of elastase-like enzyme activities than burn blisters or fresh suction blisters. The gel filtration studies using Sepharose CL-4B chromatography revealed two major peaks of SAPNA hydrolysing enzyme activity in burn blister and in bullous pemphigoid blister. The first peak eluted in void volume, and the second peak had an apparent molecular weight of 2.5 X 10(5) daltons. The results indicate that various proteinases are present in blisters fluids and that they may participate in the blister formation by degrading structural components of the basement membrane zone and the dermis.