Cathepsin D is present in human eccrine sweat and involved in the postsecretory processing of the antimicrobial peptide DCD-1L

被引:78
作者
Baechle, D
Flad, T
Cansier, A
Steffen, H
Schittek, B
Tolson, J
Herrmann, T
Dihazi, H
Beck, A
Mueller, GA
Mueller, M
Stevanovic, S
Garbe, C
Mueller, CA
Kalbacher, H
机构
[1] Univ Tubingen, Med & Nat Sci Res Ctr, D-72074 Tubingen, Germany
[2] Univ Tubingen, Sect Transplantat Immunol & Immunohematol, D-72072 Tubingen, Germany
[3] Univ Tubingen, Dept Dermatol, D-72076 Tubingen, Germany
[4] Univ Tubingen, Dept Immunol, D-72076 Tubingen, Germany
[5] Univ Hosp Gottingen, Dept Nephrol & Rheumatol, D-37075 Gottingen, Germany
[6] Univ Tubingen Hosp, Dept Internal Med 4, Div Clin Chem, D-72076 Tubingen, Germany
关键词
D O I
10.1074/jbc.M504670200
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The protein pattern of healthy human eccrine sweat was investigated and 10 major proteins were detected from which apolipoprotein D, lipophilin B, and cathepsin D ( CatD) were identified for the first time in human eccrine sweat. We focused our studies on the function of the aspartate protease CatD in sweat. In vitro digestion experiments using a specific fluorescent CatD substrate showed that CatD is enzymatically active in human sweat. To identify potential substrates of CatD in human eccrine sweat LL-37 and DCD-1L, two antimicrobial peptides present in sweat, were digested in vitro with purified CatD. LL-37 was not significantly digested by CatD, whereas DCD-1L was cleaved between Leu(44) and Asp(45) and between Leu(29) and Glu(30) almost completely. The DCD-1L-derived peptides generated in vitro by CatD were also found in vivo in human sweat as determined by surface-enhanced laser desorption/ ionization (SELDI) mass spectrometry. Furthermore, besides the CatD-processed peptides we identified additionally DCD-1L-derived peptides that are generated upon cleavage with a 1,10-phenanthroline- sensitive carboxypeptidase and an endoprotease. Taken together, proteolytic processing generates 12 DCD-1L-derived peptides. To elucidate the functional significance of postsecretory processing the antimicrobial activity of three CatD-processed DCD-1L peptides was tested. Whereas two of these peptides showed no activity against Gram-positive and Gram-negative bacteria, one DCD-1L-derived peptide showed an even higher activity against Escherichia coli than DCD-1L. Functional analysis indicated that proteolytic processing of DCD-1L by CatD in human sweat modulates the innate immune defense of human skin.
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页码:5406 / 5415
页数:10
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