Purification of Antimicrobial Peptides from Human Skin

Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 618)

Abstract

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 

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Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  1. 1.Clinical Research Unit “Cutaneous Inflammation”, Department of DermatologyUniversity Hospital Schleswig-HolsteinKielGermany

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