title: Kallikrein-related peptidase 5 functions in proteolytic processing of profilaggrin in cultured human keratinocytes.
abstract: Filaggrin protein is synthesized in the stratum granulosum of the skin and contributes to the formation of the human skin barrier. Profilaggrin is cleaved by proteolytic enzymes and converted to functional filaggrin, but its processing mechanism remains not fully elucidated. Kallikrein-related peptidase 5 (KLK5) is a major serine protease found in the skin, which is secreted from lamellar granules following its expression in the stratum granulosum and activated in the extracellular space of the stratum corneum. Here, we searched for profilaggrin-processing protease(s) by partial purification of epidermal extracts and found KLK5 as a possible candidate. We used high performance liquid chromatography coupled with electrospray tandem mass spectrometry to show that KLK5 cleaves profilaggrin. Furthermore, based on a proximity ligation assay, immunohistochemistry, and immunoelectron microscopy analysis, we reveal that KLK5 and profilaggrin co-localize in the stratum granulosum in human epidermis. KLK5 knockdown in normal cultured human epidermal keratinocytes resulted in higher levels of profilaggrin, indicating that KLK5 potentially functions in profilaggrin cleavage.
authors: Department of Dermatology, Hamamatsu University School of Medicine, 1-20-1 Handayama, Higashi-ku, Hamamatsu 431-3192, Japan. jsakabe@hama-med.ac.jp
authors: Jun-ichi
authors: J
authors: Sakabe
authors: Mami
authors: M
authors: Yamamoto
authors: Satoshi
authors: S
authors: Hirakawa
authors: Akira
authors: A
authors: Motoyama
authors: Isao
authors: I
authors: Ohta
authors: Kazuki
authors: K
authors: Tatsuno
authors: Taisuke
authors: T
authors: Ito
authors: Kenji
authors: K
authors: Kabashima
authors: Toshihiko
authors: T
authors: Hibino
authors: Yoshiki
authors: Y
authors: Tokura
mesh_terms: D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000818:Animals; D002478:Cells, Cultured; D000091344:Filaggrin Proteins; D055785:Gene Knockdown Techniques; D006801:Humans; D007381:Intermediate Filament Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D007610:Kallikreins / Q000737:chemistry / Q000502:physiology*; D015603:Keratinocytes / Q000201:enzymology; D051379:Mice; D008810:Mice, Inbred C57BL; D011499:Protein Processing, Post-Translational; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D059748:Proteolysis; D034741:RNA, Small Interfering / Q000235:genetics; D012867:Skin / Q000166:cytology / Q000201:enzymology
chemical_list: C000717249:FLG protein, human; D000091344:Filaggrin Proteins; D007381:Intermediate Filament Proteins; D034741:RNA, Small Interfering; C400147:KLK5 protein, human; D007610:Kallikreins
keywords: Differentiation; Epidermis; RNA Interference (RNAi); Serine Protease; Skin
references: Nature. 1986 Oct 30-Nov 5;323(6091):822-4
references: 2430189
references: J Invest Dermatol. 2005 Feb;124(2):360-6
references: 15675955
references: J Biol Chem. 2009 May 8;284(19):12829-36
references: 19286660
references: J Allergy Clin Immunol. 2012 Jun;129(6):1538-46.e6
references: 22409988
references: J Cell Sci. 2009 May 1;122(Pt 9):1285-94
references: 19386895
references: Mol Biol Cell. 2007 Sep;18(9):3607-19
references: 17596512
references: J Cell Biol. 2003 Nov 24;163(4):901-10
references: 14638864
references: J Invest Dermatol. 2004 May;122(5):1235-44
references: 15140227
references: Br J Dermatol. 2009 Dec;161(6):1387-90
references: 19663875
references: Annu Rev Biochem. 1988;57:593-625
references: 3052284
references: J Invest Dermatol. 2010 Sep;130(9):2286-94
references: 20445547
references: J Cell Biochem. 2010 Feb 15;109(3):487-97
references: 19960512
references: J Biol Chem. 2007 Feb 9;282(6):3640-52
references: 17158887
references: J Exp Med. 2009 May 11;206(5):1135-47
references: 19414552
references: FASEB J. 2011 May;25(5):1567-76
references: 21282207
references: J Cell Biol. 2007 Apr 9;177(1):173-82
references: 17403930
references: Arch Dermatol Res. 2004 Mar;295(10):448-52
references: 14762669
references: Biochim Biophys Acta. 1981 Jun 29;669(1):65-78
references: 6170342
references: J Biol Chem. 2011 Mar 11;286(10):8204-8212
references: 21190945
references: Methods. 2008 Jul;45(3):227-32
references: 18620061
references: J Dermatol Sci. 2006 Nov;44(2):63-72
references: 16973334
references: Proc Natl Acad Sci U S A. 1983 Feb;80(3):716-20
references: 6187003
references: Differentiation. 1983;23(3):243-9
references: 6406285
references: PLoS One. 2009;4(4):e5227
references: 19384417
references: J Invest Dermatol. 2012 May;132(5):1507-10
references: 22318384
references: J Cell Biol. 2005 Aug 1;170(3):487-96
references: 16061697
references: EMBO Mol Med. 2011 Jun;3(6):320-33
references: 21542132
references: J Invest Dermatol. 2009 Jun;129(6):1446-58
references: 19020553
references: Cell. 1982 Nov;31(1):11-24
references: 6186379
references: J Invest Dermatol. 1979 May;72(5):257-61
references: 88487
references: Biochemistry. 1984 Mar 13;23(6):1239-45
references: 6712945
references: J Biol Chem. 1978 Aug 25;253(16):5861-8
references: 670237
references: J Biol Chem. 2012 Sep 21;287(39):32825-34
references: 22825846