Antimicrobial Peptides

Volume 618 of the series Methods in Molecular Biology pp 15-30


Purification of Antimicrobial Peptides from Human Skin

  • Jens-M. SchröderAffiliated withClinical Research Unit “Cutaneous Inflammation”, Department of Dermatology, University Hospital Schleswig-Holstein Email author 

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Human skin is a rich source of human antimicrobial peptides. Its cellular source is the keratinocyte, which terminally differentiates in the uppermost parts of the skin, eventually forming the stratum corneum, the horny layer. The easy availability of human stratum corneum makes it possible to identify and characterize human antimicrobial peptides with a biochemical approach. Moreover, the availability of lesional scales of patients with psoriasis, an inflammatory skin disease, allows the identification of human-inducible peptide antibiotics, which are absent in healthy skin. With this strategy, the beta-defensins hBD-2 and hBD-3, RNase-7 as well as psoriasin/S100A7 have been discovered as human antimicrobial peptides and proteins. A detailed description of the strategies and methods is presented, which allowed a successful identification and characterization of human antimicrobial peptides. We used various HPLC techniques, combined with antimicrobial testing as read-out system. In parallel, SDS-PAGE analyses as well as electrospray ionization mass spectrometry were used for further biochemical characterization as well as purity assessment.

Key words

Antimicrobial peptides defensin RNase-7 S100A7 skin stratum corneum purification electrospray ionization mass spectrometry high-performance liquid chromatography