Use of the tape stripping technique for directly quantifying esterase activities in human stratum corneum.
The enzymes secreted in the intercellular spaces of stratum corneum (SC), the outermost layer of the epidermis, are thought to be involved in normal desquamation and skin barrier function. Their activity can barely be measured due to the difficulty in isolating enough biological material. Human SC layers were obtained from the forearm of healthy volunteers by the tape stripping technique. Assays for esterase activities were carried out in specially designed plates which contained the SC blotted on tape strips, using various fluorescent methylumbelliferone acyl esters as substrates. Triacylglycerol hydrolase activities were also studied by this method. By using radiolabeled triolein and fluorescent 4-methylumbelliferyl 7-oleate as substrates, true lipase activities could be detected and quantitated in SC at pH 5.5 and 7.5. These activities were shown to be strongly inhibited by tetrahydrolipstatin while this was not the case with 4-methylumbelliferyl 7-heptanoate. The method described here combines the painless tape stripping technique with a sensitive plate assay analysis. Since the whole process needs little manipulation, this method can permit rapid quantitation of multiple enzyme activities from a single strip. Therefore, it will permit the study of the involvement of enzyme activities in epidermis aging and skin pathologies.
Beisson
F
F
Laboratoire de Lipolyse Enzymatique (UPR 9025 du CNRS), Institut de Biologie Structurale et Microbiologie du CNRS et de l'Université de la Méditerranée (FR 2248), 31 chemin Joseph Aiguier, 13402 Marseille Cedex 20, France.
Aoubala
M
M
Marull
S
S
Moustacas-Gardies
A M
AM
Voultoury
R
R
Verger
R
R
Arondel
V
V
D004817:Epidermis / Q000201:enzymology*; D004950:Esterases / Q000032:analysis*; D005542:Forearm; D006801:Humans; D008722:Methods
D004950:Esterases