SKALP/elafin is an inducible proteinase inhibitor in human epidermal keratinocytes.
Skin-derived antileukoproteinase (SKALP), otherwise known as elafin, is a recently discovered epidermal proteinase inhibitor with specificity for polymorphonuclear leukocyte (PMN)-derived elastase and proteinase-3; in addition to the proteinase-inhibiting domain, SKALP contains several transglutaminase substrate motifs. SKALP is virtually absent in normal human epidermis but is found in a number of inflammatory skin diseases, including psoriasis. Here we report the induction and processing of SKALP in vivo and in vitro. SKALP expression in vivo could be demonstrated following injury in normal human epidermis, using histology, western blotting, northern blotting and a functional assay. In vitro, SKALP expression was studied in conventional submerged keratinocyte culture systems and in keratinocytes cultured in an air-liquid interface model. Induction of SKALP activity in epidermis could be measured as early as 16 hours after skin injury; immunohistological examination showed that SKALP expression was confined to the outer layers of the stratum spinosum and the stratum granulosum. Northern blot analysis revealed a 0.8 kb transcript, both in vivo (psoriatic skin, injured skin) and in vitro (cultured keratinocytes). Western blot analysis showed that the major SKALP form in vivo was a low molecular mass fragment, containing the antiproteinase domain. In all cultures that were positive for SKALP, larger (8-10 kDa) forms of SKALP, containing the N-terminal transglutaminase substrate motifs in addition to the antiproteinase domain, were found. SKALP expression in cultured cells was found to be dependent on the system used. In a submerged culture system, SKALP could be induced by fetal calf serum.(ABSTRACT TRUNCATED AT 250 WORDS)
Alkemade
J A
JA
Department of Dermatology, University Hospital Nijmegen, The Netherlands.
Molhuizen
H O
HO
Ponec
M
M
Kempenaar
J A
JA
Zeeuwen
P L
PL
de Jongh
G J
GJ
van Vlijmen-Willems
I M
IM
van Erp
P E
PE
van de Kerkhof
P C
PC
Schalkwijk
J
J
D000388:Air; D000595:Amino Acid Sequence; D002118:Calcium / Q000494:pharmacology; D046508:Culture Techniques / Q000379:methods; D000078404:Epidermal Cells; D004817:Epidermis / Q000378:metabolism*; D005786:Gene Expression Regulation; D006801:Humans; D015603:Keratinocytes / Q000378:metabolism*; D008969:Molecular Sequence Data; D053491:Proteinase Inhibitory Proteins, Secretory; D011506:Proteins; D011565:Psoriasis / Q000378:metabolism*; D012333:RNA, Messenger / Q000302:isolation & purification; D015842:Serine Proteinase Inhibitors / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D012867:Skin / Q000033:anatomy & histology / Q000254:growth & development; D014018:Tissue Distribution
D053491:Proteinase Inhibitory Proteins, Secretory; D011506:Proteins; D012333:RNA, Messenger; D015842:Serine Proteinase Inhibitors; D002118:Calcium