Cloning and characterization of hurpin (protease inhibitor 13): A new skin-specific, UV-repressible serine proteinase inhibitor of the ovalbumin serpin family

Epidermal keratinocytes are the primary target of the midrange ultraviolet part (UVB, 280-320 nm) of terrestrial sunlight. Analysis of the resulting U V response at the transcriptional level by differential display PCR identif ied a formerly unrecognized large group of repressed genes. Among those UV- repressible genes, a novel serine proteinase inhibitor (serpin) termed hurp in (HaCaT UV-repressible serpin) has been identified. The isolated full-len gth cDNAs harbour a 1176 bp open reading frame encoding a potential protein with 391 amino acid residues and a predicted molecular mass of similar to 44 kDa. The novel serpin has nearly 59% amino acid identity with the squamo us cell carcinoma antigen 1 (SCCA1) and squamous cell carcinoma antigen 2 ( SCCA2). In addition, it displays all of the structural features unique to t he ovalbumin family of serpins (ov-serpins). The amino acid sequence of the hinge region in the reactive site loop suggests that hurpin has the potent ial for protease inhibition. The putative reactive center P-1-P-1, residues were identified as Thr356-Ser357 by alignment with other ov-serpins. The p hysiological target protease is unknown and the in vitro translated hurpin does not form SDS-stable complexes with a variety of known serine proteases . Expression of hurpin is restricted to epidermal cells where two distinct transcripts of 3.0 and 3.4 kb are detectable. Furthermore, expression of hu rpin appears to be related to the activation or proliferation state of kera tinocytes, since hurpin transcripts are more abundant in immortalized kerat inocytes (HaCaT) and in cultured normal human keratinocytes, compared to th e expression in normal skin. Moreover, in psoriasis, a skin disease charact erized by hyperproliferation of keratinocytes and responsive to therapeutic UV irradiation, overexpression of hurpin is noted in psoriatic skin lesion s compared to non-lesional skin. (C) 1999 Academic Press.